782 results
Search Results
2. Enhanced Intracellular Reactive Oxygen Species by Photodynamic Therapy Effectively Promotes Chemoresistant Cell Death
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Xiaolin Xu, Chenglong Wang, Peipei Zhang, Xuzhu Gao, Wencai Guan, Fanchen Wang, Xin Li, Jia Yuan, Hongjing Dou, and Guoxiong Xu
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Mice, Inbred BALB C ,Cell Death ,malignant tumor ,oxidative phosphorylation ,ROS ,Cell Biology ,glycolysis ,Xenograft Model Antitumor Assays ,Applied Microbiology and Biotechnology ,Mice ,Photochemotherapy ,photodynamic therapy ,Drug Resistance, Neoplasm ,Cell Line, Tumor ,Animals ,Humans ,Female ,Reactive Oxygen Species ,Molecular Biology ,Chemoresistance ,Ecology, Evolution, Behavior and Systematics ,Research Paper ,Developmental Biology - Abstract
Anti-cancer chemo-drugs can cause a rapid elevation of intracellular reactive oxygen species (ROS) levels. An imbalance in ROS production and elimination systems leads to cancer cell resistance to chemotherapy. This study aimed to evaluate the mechanism and effect of ROS on multidrug resistance in various human chemoresistant cancer cells by detecting the changes in the amount of ROS, the expression of ROS-related and glycolysis-related genes, and cell death. We found that ROS was decreased while oxidative phosphorylation was increased in chemoresistant cells. We verified that the chemoresistance of cancer cells was achieved in two ways. First, chemoresistant cells preferred oxidative phosphorylation instead of anaerobic glycolysis for energy generation, which increased ATPase activity and produced much more ATP to provide energy. Second, ROS-scavenging systems were enhanced in chemoresistant cancer cells, which in turn decreased ROS amount and thus inhibited chemo-induced cell death. Our in vitro and in vivo photodynamic therapy further demonstrated that elevated ROS production efficiently inhibited chemo-drug resistance and promoted chemoresistant cell death. Taken together, targeting ROS systems has a great potential to treat cancer patients with chemoresistance.
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- 2022
3. Phyto‐fabricated silver nanoparticles inducing microbial cell death via reactive oxygen species‐mediated membrane damage
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Ch. V. Ramana Devi and L. Srinivas Naik
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Silver ,Biocompatibility ,Metal Nanoparticles ,Microbial Sensitivity Tests ,Bacillus subtilis ,Silver nanoparticle ,Enterobacteriaceae ,Momordica dioica ,Electrical and Electronic Engineering ,Cell Death ,biology ,Plant Extracts ,Chemistry ,Biofilm ,Antimicrobial ,biology.organism_classification ,Anti-Bacterial Agents ,Electronic, Optical and Magnetic Materials ,Original Research Paper ,Membrane ,Nucleic acid ,Reactive Oxygen Species ,Original Research Papers ,TP248.13-248.65 ,Biotechnology ,Nuclear chemistry - Abstract
Eco‐friendly synthesis of the silver nanoparticles (AgNPs) has a number of advantages like simplicity, biocompatibility, low toxicity in nature over their physical and chemical methods. In the present study, the authors report biosynthesized AgNPs using the root extract of the perennial plant ‘Spiny gourd’ (Momordica dioica) and investigated their anti‐bacterial application with mechanistic approaches. Different biophysical techniques such as UV‐Vis spectroscopy, FTIR, XRD, TEM, SAED, and DLS were employed for AgNPs characterization. The synthesized AgNPs were polydispersed, crystalline in nature, with anionic surface (−22.3 mV), spherical in shape with an average size of 13.2 nm. In addition, the AgNPs were stable in room temperature and in different biological buffers. The anti‐bacterial activities of AgNPs were studied with respect to the pathogens such as Bacillus subtilis, Staphylococcus aureus (Gram‐positive), Pseudomonas aeruginosa, Escherichia coli, Klebsiella planticola (Gram‐negative), and Candida albicans. Also, mechanistic studies of AgNPs such as protein leakage assay, nucleic acid leakage assay, ATP leakage assay, ROS accumulation, determination of biofilm degrading activity, measurement of potassium, showing that the synthesized AgNPs are capable of containing a potential application in the antimicrobial therapeutic agents and the pharmaceutical industry.
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- 2021
4. Investigating the in vitro photothermal effect of green synthesized apigenin‐coated gold nanoparticle on colorectal carcinoma
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Seyed Mohammad Amini, Ali Neshasteh-Riz, Elham Mohammadi, Ali Shakeri-Zadeh, Shaghayegh Askarian-Amiri, and Yaser Azizi
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Programmed cell death ,Metal Nanoparticles ,Nanoparticle ,Mice ,chemistry.chemical_compound ,Animals ,Apigenin ,Electrical and Electronic Engineering ,Cell Death ,Chemistry ,Photothermal effect ,Photothermal therapy ,bacterial infections and mycoses ,equipment and supplies ,Combinatorial chemistry ,Electronic, Optical and Magnetic Materials ,Original Research Paper ,Colloidal gold ,Apoptosis ,bacteria ,Chemical binding ,Gold ,Colorectal Neoplasms ,Original Research Papers ,TP248.13-248.65 ,Biotechnology - Abstract
Applying toxic chemical to the synthesis of stable gold nanoparticles is one of the limitations of gold nanoparticles for therapeutic applications such as photothermal therapy. Plant compounds such as apigenin (API) with therapeutic potential can be applied in the synthesis of gold nanoparticles. API‐coated gold nanoparticles (Api@AuNPs) with an average size of 19.1 nm and a surface charge of −4.3 mV have been synthesized by a simple and efficient technique. The stability of Api@AuNPs in the biological environment was verified through UV‐Vis spectroscopy. Based on Raman and FTIR spectroscopy analysis, chemical binding of API on the surface of Api@AuNPs through hydroxyl and carbonyl functional groups was found to be the main reason for the stability of the Api@AuNPs in comparison with citrate‐coated gold nanoparticles (Cit@AuNPs). The synthesized Api@AuNPs do not cause major toxic effects up to 128 ppm. Api@AuNP‐mediated photothermal therapy leads to the indiscriminate eradication of almost half of both mouse fibroblastic (L929) and colorectal cancer (CT26) cells. Flow‐cytometry analysis revealed that the cell death mechanism is mainly apoptosis. In the apoptosis triggered cell death in photothermal treatment, Api@AuNPs are preferred over commonly used gold nanoparticles in photothermal treatments which mostly trigger the necrosis cell death pathway.
- Published
- 2021
5. Celecoxib prevents tumor necrosis factor-α (TNF-α)-induced cellular senescence in human chondrocytes
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Qunli Wang, Qi Chen, Jie Sui, Yuanyuan Tu, Xiang Guo, and Feng Li
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Cyclin-Dependent Kinase Inhibitor p21 ,chondrocytes ,Bioengineering ,Ataxia Telangiectasia Mutated Proteins ,Resting Phase, Cell Cycle ,Applied Microbiology and Biotechnology ,Cell Line ,Humans ,Phosphorylation ,Telomerase ,Cellular Senescence ,Cell Death ,Tumor Necrosis Factor-alpha ,G1 Phase ,Cell Cycle Checkpoints ,General Medicine ,Checkpoint Kinase 2 ,osteoarthritis ,Celecoxib ,cell senescence ,Tumor Suppressor Protein p53 ,Reactive Oxygen Species ,TP248.13-248.65 ,Research Article ,Research Paper ,Biotechnology - Abstract
Osteoarthritis (OA) is a cartilage degenerative disease commonly observed in the elderly population and significantly impacts the normal life of OA patients. It has been reported that the development of pathological cell senescence in chondrocytes is involved in the pathogenesis of OA. Celecoxib is a common non-steroidal anti-inflammatory drug, and it has been recently reported to exert therapeutic effects on OA. However, its underlying mechanism is still unclear. The present study intends to explore its mechanism and provide fundamental evidence for the application of Celecoxib in the treatment of clinical OA. Tumor necrosis factor-α (TNF-α) was utilized to establish an in vitro model of chondrocytes senescence. The elevated reactive oxygen species (ROS) generation, increased cell cycle arrest in G0/G1 phase, reduced telomerase activity, and upregulated senescence-associatedβ-galactosidase (SA-β-Gal) staining were all observed in TNF-α-treated chondrocytes, which were then dramatically reversed by 10 and 20 μM Celecoxib. In addition, the upregulated DNA damage biomarkers, p-ATM, and p-CHK2, observed in TNF-α-treated chondrocytes were significantly downregulated by 10 and 20 μM Celecoxib. Lastly, the expression level of p21 and p53 was greatly elevated in chondrocytes by stimulation with TNF-α which was then pronouncedly repressed by treatment with Celecoxib. Taken together, our data reveal that Celecoxib ameliorated TNF-α-induced cellular senescence in human chondrocytes.
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- 2021
6. Synthesis and Characterization of Telmisartan‐Derived Cell Death Modulators to Circumvent Imatinib Resistance in Chronic Myeloid Leukemia
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Anna M. Schoepf, Ronald Gust, Verena Hohn, Petra Obexer, Florina Veider, and Stefan Salcher
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Programmed cell death ,Indoles ,Cell ,Carbazoles ,01 natural sciences ,Biochemistry ,peroxisome proliferator-activated receptor gamma ,chronic myeloid leukemia ,Cancer stem cell ,Leukemia, Myelogenous, Chronic, BCR-ABL Positive ,hemic and lymphatic diseases ,Chlorocebus aethiops ,Drug Discovery ,medicine ,Animals ,Humans ,Telmisartan ,General Pharmacology, Toxicology and Pharmaceutics ,Receptor ,Pharmacology ,Full Paper ,Cell Death ,Molecular Structure ,010405 organic chemistry ,Chemistry ,structure-activity relationship ,Organic Chemistry ,Myeloid leukemia ,Imatinib ,Full Papers ,0104 chemical sciences ,PPAR gamma ,010404 medicinal & biomolecular chemistry ,medicine.anatomical_structure ,Drug Resistance, Neoplasm ,sensitizers ,COS Cells ,imatinib resistance ,Imatinib Mesylate ,Cancer research ,Molecular Medicine ,K562 Cells ,K562 cells ,medicine.drug - Abstract
New strategies to eradicate cancer stem cells in chronic myeloid leukemia (CML) include a combination of imatinib with peroxisome proliferator‐activated receptor gamma (PPARγ) ligands. Recently, we identified the partial PPARγ agonist telmisartan as effective sensitizer of resistant K562 CML cells to imatinib treatment. Here, the importance of the heterocyclic core on the cell death‐modulating effects of the telmisartan‐derived lead 4′‐((2‐propyl‐1H‐benzo[d]imidazol‐1‐yl)methyl)‐[1,1′‐biphenyl]‐2‐carboxylic acid (3 b) was investigated. Inspired by the pharmacodynamics of HYL‐6d and the selective PPARγ ligand VSP‐51, the benzimidazole was replaced by a carbazole or an indole core. The results indicate no correlation between PPARγ activation and sensitization of resistant CML cells to imatinib. The 2‐COOH derivatives of the carbazoles or indoles achieved low activity at PPARγ, while the benzimidazoles showed 60‐100 % activation. Among the 2‐CO2CH3 derivatives, only the ester of the lead (2 b) slightly activated PPARγ. Sensitizing effects were further observed for this non‐cytotoxic 2 b (80 % cell death), and to a lesser extent for the lead 3 b or the 5‐Br‐substituted ester of the benzimidazoles (5 b)., Heterocyclic compounds with carbazole, benzimidazole, or indole core were synthesized as sensitizers to imatinib treatment in resistant chronic myelogenous leukemia cells. The benzimidazole derivatives were clearly more potent as cell death modulators than the respective carbazoles or indoles. These results are important for the further development of effective sensitizers to circumvent tyrosine kinase inhibitor resistance.
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- 2020
7. Identification of novel hepatitis B virus therapeutic vaccine candidates derived from polymerase protein
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Jinming Wu, Yilun Xu, Juzeng Zheng, Yang Liu, Xianfan Lin, Ziqiang Xia, Sisi Jin, and Zhanfan Ou
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Gene Expression Regulation, Viral ,Hepatitis B virus ,Aging ,Peptide binding ,DNA-Directed DNA Polymerase ,Biology ,medicine.disease_cause ,Peripheral blood mononuclear cell ,Fluorescence ,Epitope ,Epitopes ,Interferon-gamma ,Viral Proteins ,Immune system ,Cell Line, Tumor ,vaccine ,Gene expression ,medicine ,Humans ,Amino Acid Sequence ,Polymerase ,epitope ,Cell Death ,Viral Vaccines ,bioinformatics ,Cell Biology ,Virology ,digestive system diseases ,Molecular Docking Simulation ,polymerase ,Viral replication ,biology.protein ,Peptides ,Protein Binding ,Research Paper - Abstract
Hepatitis B virus (HBV) infection is a worldwide health problem with high morbidity and mortality rates. The therapeutic vaccine is a promising method of treatment, and HBV polymerase plays a vital role in viral replication. Therefore, a therapeutic vaccine that binds to HBV DNA polymerase may control HBV infection. We predicted and selected epitopes of polymerase using online databases and analysis software. We then performed molecular docking and peptide binding assays to evaluate the binding energies and affinities between polymerase epitopes and the HLA-A0201 molecule. Finally, we induced T cells from the peripheral blood mononuclear cells (PBMCs) of healthy donors using each epitope and quantified the functions of epitope-specific T cells by IFN-γELISPOT assay, T2 cell cytotoxicity assay, HepG2.2.15 cell cytotoxicity assay and HBV gene expression assays. Four epitopes (RVTGGVFLV, GLLGFAAPF, LLDDEAGPL and YMDDVVLGA) had low binding energy and two epitopes (RVTGGVFLV and GLLGFAAPF) had a high binding affinity. The T cells stimulated by two epitopes (GLLGFAAPF and HLYSHPIIL) had a greater ability to induce immune response and suppress HBV. The HBV DNA polymerase epitopes identified in this study are promising targets for designing an epitope-based therapeutic vaccine against HBV.
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- 2021
8. Lipotoxicity reduces DDX58/Rig-1 expression and activity leading to impaired autophagy and cell death
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Raymond G. Franks, Dividutta Das, Alyssa M. Brown, Michael Hayward, Joseph T. Nickels, Jessie Lee Cunningham, and Karla K. Frietze
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0301 basic medicine ,Programmed cell death ,Palmitic Acid ,Biology ,Mice ,03 medical and health sciences ,Sequestosome 1 ,Non-alcoholic Fatty Liver Disease ,Enhancer binding ,Sequestosome-1 Protein ,Nonalcoholic fatty liver disease ,Autophagy ,CEBPB ,medicine ,Animals ,education ,Molecular Biology ,Inflammation ,education.field_of_study ,Cell Death ,030102 biochemistry & molecular biology ,Cell Biology ,medicine.disease ,030104 developmental biology ,Lipotoxicity ,Saturated fatty acid ,Cancer research ,Research Paper - Abstract
Nonalcoholic fatty liver disease (NAFLD) is the leading cause of chronic liver disease globally. NAFLD is a consequence of fat accumulation in the liver leading to lipotoxicity. Increasing evidence has demonstrated the critical role of autophagy in NAFLD. This study uncovers the unexpected role of immune surveillance protein DDX58/Rig-1 (DExD/H box helicase 58) in activating macroautophagy/autophagy and protecting from lipotoxicity associated with NAFLD. Here we show for the first time that DDX58 protein is significantly reduced in nonalcoholic steatohepatitis (NASH) mouse model, an aggressive form of NAFLD characterized by inflammation and fibrosis of the liver. In addition to decreased expression of DDX58, we found that DDX58 activity can be attenuated by treatments with palmitic acid (PA), a saturated fatty acid. To investigate whether PA inhibition of DDX58 is harmful to the cell, we characterized DDX58 function in hepatocytes when exposed to high doses of PA in the presence and/or absence of DDX58. We show that siRNA knockdown of DDX58 promotes apoptosis. Importantly, we show that stable overexpression of DDX58 is protective against toxic levels of PA and stimulates autophagy. This study begins to demonstrate the regulation of the autophagy receptor protein SQSTM1/p62 through DDX58. DDX58 expression directly influences SQSTM1 mRNA and protein levels. This work proposes a model in which activating DDX58 increases an autophagic response and this aids in clearing toxic lipid inclusion bodies, which leads to inflammation and apoptosis. Activating a DDX58-induced autophagy response may be a strategy for treating NAFLD. Abbreviations:5ʹpppdsRNA: 5ʹ triphosphate double-stranded RNA; CDAHFD: choline-deficient, L-amino acid defined high-fat diet; CEBPB: CCAAT/enhancer binding protein (C/EBP), beta; CQ: chloroquine; DDX58/retinoic acid inducible gene 1/Rig-1: DExD/H box helicase 58; h: hours; IFIH1/MDA5: interferon induced with helicase C domain 1; IFNB/IFN-β: interferon beta 1, fibroblast; KO: knockout; MAVS: mitochondrial antiviral signaling protein; NAFLD: nonalcoholic fatty liver disease; NASH: nonalcoholic steatohepatitis; NFKB/NF-κB: nuclear factor of kappa light polypeptide gene enhancer in B cells; PA: palmitic acid; poly:IC: polyinosinic:polycytidylic acid; PRR: pattern recognition receptors; PSR: picrosirus red; RAP: rapamycin; RLR: RIG-I-like receptor; SQSTM1/p62: sequestosome 1; STING1: stimulator of interferon response cGAMP interactor 1; TBK1: TANK-binding kinase 1.
- Published
- 2021
9. Acetyl oxygen benzoate engeletin ester promotes KLF4 degradation leading to the attenuation of pulmonary fibrosis via inhibiting TGFβ1–smad/p38MAPK–lnc865/lnc556–miR-29b-2-5p–STAT3 signal pathway
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Youlei Wang, Ruiqiong Li, Xiaodong Song, Ke Shen, Changjun Lv, Bo Liu, Xiaoli Zhang, Guiwu Qu, Rongrong Li, and Minge Li
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STAT3 Transcription Factor ,Aging ,Flavonols ,Kruppel-Like Transcription Factors ,Engeletin ,Smad Proteins ,SMAD ,Bleomycin ,Models, Biological ,p38 Mitogen-Activated Protein Kinases ,Cell Line ,STAT3 ,Transforming Growth Factor beta1 ,Kruppel-Like Factor 4 ,chemistry.chemical_compound ,lncRNA ,Pulmonary fibrosis ,medicine ,Animals ,Glycosides ,Fibroblast ,miRNA ,pulmonary fibrosis ,Cell Death ,biology ,Activator (genetics) ,Cell Biology ,medicine.disease ,KLF4 ,Mice, Inbred C57BL ,MicroRNAs ,medicine.anatomical_structure ,chemistry ,Proteolysis ,Cancer research ,biology.protein ,RNA, Long Noncoding ,Research Paper ,Signal Transduction - Abstract
Pulmonary fibrosis is a common pulmonary interstitial disease of pathogenesis without effective drugs for treatment. Therefore, discovering new and effective drugs is urgently needed. In the present study, we prepared a novel compound named acetyl oxygen benzoate engeletin ester (AOBEE), investigated its effect on experimental pulmonary fibrosis, and proposed a long non-coding RNA (lncRNA)-mediated mechanism of its action. Bleomycin-induced pulmonary fibrosis in mice exhibited that AOBEE improved forced vital capacity (FVC) and alveolar structure and inhibited α-SMA, vimentin, and collagen expression. TGFβ1-stimulated fibroblast L929 cells showed that AOBEE reduced these fibrotic proteins expression and inhibited the activated-fibroblast proliferation and migration. Whole transcriptome sequencing was performed to screen out lncRNA-lnc865 and lnc556 with high expression under bleomycin treatment, but AOBEE caused a considerable decrease in lnc865 and lnc556. Mechanistic study elucidated that AOBEE alleviated pulmonary fibrosis through lnc865- and lnc556-mediated mechanism, in which both lnc865 and lnc556 sponged miR-29b-2-5p to target signal transducer and activator of transcription 3 (STAT3). Further signal pathway inhibitors and the Cignal Finder 45-pathway reporter array illustrated that the up- and downstream pathways were TGFβ1-smad2/3 and p38MAPK, and Krüppel-like factor 4 (KLF4), respectively. In conclusion, AOBEE promoted KLF4 degradation leading to the attenuation of pulmonary fibrosis by inhibiting TGFβ1-smad/p38MAPK-lnc865/lnc556-miR-29b-2-5p-STAT3 signal pathway. We hope this work will provide valuable information to design new drugs and therapeutic targets of lncRNAs for pulmonary fibrosis treatment.
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- 2021
10. Exploration of p53 plus interferon-beta gene transfer for the sensitization of human colorectal cancer cell lines to cell death
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Rodrigo Esaki Tamura, Samir Andrade Mendonça, Fernanda Antunes, Bryan E. Strauss, Daniela B Zanatta, Aline Hunger, and Paulo Roberto Del Valle
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0301 basic medicine ,Cancer Research ,Programmed cell death ,Colorectal cancer ,medicine.medical_treatment ,Genetic enhancement ,Apoptosis ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Humans ,Sensitization ,Pharmacology ,Chemotherapy ,Cell Death ,business.industry ,Gene Transfer Techniques ,Interferon-beta ,HCT116 Cells ,medicine.disease ,030104 developmental biology ,medicine.anatomical_structure ,Oncology ,Cell culture ,030220 oncology & carcinogenesis ,Cancer research ,Molecular Medicine ,Immunogenic cell death ,Tumor Suppressor Protein p53 ,Colorectal Neoplasms ,business ,Research Paper - Abstract
While treatments for colorectal cancer continue to improve, some 50% of patients succumb within 5 years, pointing to the need for additional therapeutic options. We have developed a modified non-replicating adenoviral vector for gene transfer, called AdRGD-PG, which offers improved levels of transduction and transgene expression. Here, we employ the p53-responsive PG promoter to drive expression of p53 or human interferon-β (hIFNβ) in human colorectal cancer cell lines HCT116(wt) (wtp53), HCT116(−/-) (p53 deficient) and HT29 (mutant p53). The HCT116 cell lines were both easily killed with p53 gene transfer, while combined p53 and hIFNβ cooperated for the induction of HT29 cell death and emission of immunogenic cell death (ICD) markers. Elevated annexinV staining and caspase 3/7 activity point to cell death by a mechanism consistent with apoptosis. P53 gene transfer alone or in combination with hIFNβ sensitized all cell lines to chemotherapy, permitting the application of low drug doses while still achieving significant loss of viability. While endogenous p53 status was not sufficient to predict response to treatment, combined p53 and hIFNβ provided an additive effect in HT29 cells. We propose that this approach may prove effective for the treatment of colorectal cancer, permitting the use of limited drug doses.
- Published
- 2021
11. CPI-1189 protects neuronal cells from oxygen glucose deprivation/re-oxygenation-induced oxidative injury and cell death
- Author
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Chengliang Yang, Yueli Zhan, Chengrui Li, Jianhong Sun, and Yong-Jun Li
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Aging ,Programmed cell death ,oxidative injury ,p38 mitogen-activated protein kinases ,Apoptosis ,Neuroprotection ,Cell Line ,Lipid peroxidation ,chemistry.chemical_compound ,Animals ,Neurons ,chemistry.chemical_classification ,Reactive oxygen species ,Cell Death ,oxygen glucose deprivation/re-oxygenation ,Depolarization ,Cell Biology ,Glutathione ,CPI-1189 ,Cell biology ,Oxygen ,Oxidative Stress ,Glucose ,nervous system ,chemistry ,Butanes ,Tumor necrosis factor alpha ,Lipid Peroxidation ,signaling ,Reactive Oxygen Species ,Research Paper - Abstract
Oxygen glucose deprivation (OGD)/re-oxygenation (OGDR) induces profound oxidative injury and neuronal cell death. It mimics ischemia-reperfusion neuronal injury. CPI-1189 is a novel tumor necrosis factor alpha-inhibiting compound with potential neuroprotective function. Here in SH-SY5Y neuronal cells and primary murine cortical neurons, CPI-1189 pretreatment potently inhibited OGDR-induced viability reduction and cell death. In OGDR-stimulated neuronal cells, p38 phosphorylation was blocked by CPI-1189. In addition, CPI-1189 alleviated OGDR-induced reactive oxygen species production, lipid peroxidation, and glutathione consumption. OGDR-induced neuronal cell apoptosis was also inhibited by CPI-1189 pretreatment. Furthermore, in SH-SY5Y cells and cortical neurons, CPI-1189 alleviated OGDR-induced programmed necrosis by inhibiting mitochondrial p53-cyclophilin D-adenine nucleotide translocase 1 association, mitochondrial depolarization, and lactate dehydrogenase release to the medium. In summary, CPI-1189 potently inhibited OGDR-induced oxidative injury and neuronal cell death.
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- 2021
12. Triptolide improves neurobehavioral functions, inflammation, and oxidative stress in rats under deep hypothermic circulatory arrest
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Jian-Feng Liu, Zeng-Chun Wang, Hua Cao, Qiang Chen, and Yu-Qing Lei
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Male ,Aging ,deep hypothermia circulatory arrest (DHCA) ,Spatial Learning ,Morris water navigation task ,Pharmacology ,neurotrophins ,medicine.disease_cause ,Neuroprotection ,neuroinflammation ,chemistry.chemical_compound ,medicine ,Animals ,Nerve Growth Factors ,Neuroinflammation ,Inflammation ,TUNEL assay ,Behavior, Animal ,Cell Death ,business.industry ,Brain ,Cell Biology ,Phenanthrenes ,Triptolide ,Hypothermia ,Rats ,Circulatory Arrest, Deep Hypothermia Induced ,Oxidative Stress ,Memory, Short-Term ,Neuroprotective Agents ,chemistry ,triptolide ,Deep hypothermic circulatory arrest ,Epoxy Compounds ,Microglia ,Diterpenes ,medicine.symptom ,business ,Oxidative stress ,Research Paper - Abstract
This study investigated the neuroprotective effects of triptolide (TPL) in a rat model of cardiopulmonary bypass with deep hypothermia circulatory arrest (DHCA). Rats were randomly divided into six groups: control, sham, DHCA, and DHCA + TPL (100, 200, 300 μg/kg). Neurobehavioral functions were measured using the elevated plus-maze, Y-maze, and Morris water maze tests. Levels of inflammatory cytokines, oxidative stress indices, and brain neurotrophins were measured by ELISA. Microglial activation and cell death was measured by immunofluorescence staining and TUNEL assay, respectively. Finally, activation of the Nrf2 pathway and NF-κB were detected by western blot. The elevated plus-maze, Y-maze, and Morris water maze tests all showed that TPL mitigated anxiety-like behavior, working memory, spatial learning, and memory in DHCA rats. TPL inhibited inflammatory responses and oxidative stress, as well as increased brain neurotrophin levels in DHCA rats. Moreover, TPL attenuated microglia activation and cell death in DHCA rats. Finally, TPL activated the Nrf2 pathway and inhibited NF-κB activity in DHCA rats. These results demonstrated that TPL improved neurobehavioral functions, neuroinflammation, and oxidative stress in DHCA rats, which may be associated with the Nrf2 and NF-κB pathways.
- Published
- 2021
13. Orthogonal targeting of osteoclasts and myeloma cells for radionuclide stimulated dynamic therapy induces multidimensional cell death pathways
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Lynne Marsala, Gregory M. Lanza, Julie L. Prior, Alexander Zheleznyak, Xiaoxia Yang, Baogang Xu, Samuel Achilefu, Steven Fletcher, Grace Cui, Francesca Fontana, and Matthew Mixdorf
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Programmed cell death ,bone marrow ,Osteolysis ,photosensitizer ,Osteoclasts ,Medicine (miscellaneous) ,Apoptosis ,Bone Neoplasms ,Bone and Bones ,Theranostic Nanomedicine ,Mice ,Cerenkov radiation ,Fluorodeoxyglucose F18 ,In vivo ,Cell Line, Tumor ,Bone cell ,Organometallic Compounds ,Tumor Microenvironment ,medicine ,Animals ,Humans ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,orthogonal drug delivery ,Radioisotopes ,Tumor microenvironment ,nanomicelles ,Cell Death ,Chemistry ,medicine.disease ,Xenograft Model Antitumor Assays ,multiple myeloma ,medicine.anatomical_structure ,Tumor progression ,Cancer research ,Bone marrow ,Radiopharmaceuticals ,Reactive Oxygen Species ,Integrin alpha Chains ,Research Paper ,Signal Transduction - Abstract
Rationale: Multiple myeloma (MM) is a multifocal malignancy of bone marrow plasma cells, characterized by vicious cycles of remission and relapse that eventually culminate in death. The disease remains mostly incurable largely due to the complex interactions between the bone microenvironment (BME) and MM cells (MMC). In the “vicious cycle” of bone disease, abnormal activation of osteoclasts (OCs) by MMC causes severe osteolysis, promotes immune evasion, and stimulates the growth of MMC. Disrupting these cancer-stroma interactions would enhance treatment response. Methods: To disrupt this cycle, we orthogonally targeted nanomicelles (NM) loaded with non-therapeutic doses of a photosensitizer, titanocene (TC), to VLA-4 (α4ß1, CD49d/CD29) expressing MMC (MM1.S) and αvß3 (CD51/CD61) expressing OC. Concurrently, a non-lethal dose of a radiopharmaceutical, 18F-fluorodeoxyglucose ([18F]FDG) administered systemically interacted with TC (radionuclide stimulated therapy, RaST) to generate cytotoxic reactive oxygen species (ROS). The in vitro and in vivo effects of RaST were characterized in MM1.S cell line, as well as in xenograft and isograft MM animal models. Results: Our data revealed that RaST induced non-enzymatic hydroperoxidation of cellular lipids culminating in mitochondrial dysfunction, DNA fragmentation, and caspase-dependent apoptosis of MMC using VLA-4 avid TC-NMs. RaST upregulated the expression of BAX, Bcl-2, and p53, highlighting the induction of apoptosis via the BAK-independent pathway. The enhancement of multicopper oxidase enzyme F5 expression, which inhibits lipid hydroperoxidation and Fenton reaction, was not sufficient to overcome RaST-induced increase in the accumulation of irreversible function-perturbing α,ß-aldehydes that exerted significant and long-lasting damage to both DNA and proteins. In vivo, either VLA-4-TC-NM or αvß3-TC-NMs RaST induced a significant therapeutic effect on immunocompromised but not immunocompetent MM-bearing mouse models. Combined treatment with both VLA-4-TC-NM and αvß3-TC-NMs synergistically inhibited osteolysis, reduced tumor burden, and prevented rapid relapse in both in vivo models of MM. Conclusions: By targeting MM and bone cells simultaneously, combination RaST suppressed MM disease progression through a multi-prong action on the vicious cycle of bone cancer. Instead of using the standard multidrug approach, our work reveals a unique photophysical treatment paradigm that uses nontoxic doses of a single light-sensitive drug directed orthogonally to cancer and bone cells, followed by radionuclide-stimulated generation of ROS to inhibit tumor progression and minimize osteolysis in both immunocompetent murine and immunocompromised human MM models.
- Published
- 2021
14. Conditioned medium from adipose-derived stem cells attenuates ischemia/reperfusion-induced cardiac injury through the microRNA-221/222/PUMA/ETS-1 pathway
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Shu-Rung Lin, Chiang-Wen Lee, Yu-Chen Chen, Chi-Ming Pu, I-Ta Lee, Jaw-Shiun Tsai, Tzu-Lin Lee, Shu-Wha Lin, Yuh-Lien Chen, and Tsai-Chun Lai
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Male ,0301 basic medicine ,p38 mitogen-activated protein kinases ,Myocardial Infarction ,Medicine (miscellaneous) ,Apoptosis ,Myocardial Reperfusion Injury ,Ischemia/reperfusion injury ,Proto-Oncogene Protein c-ets-1 ,Andrology ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Reperfusion therapy ,Annexin ,Fibrosis ,Puma ,Animals ,Medicine ,Myocytes, Cardiac ,p53 upregulated modulator of apoptosis ,ADSC-CM ,miR-221/222 ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,Mice, Knockout ,TUNEL assay ,Cell Death ,biology ,business.industry ,Myocardium ,Stem Cells ,Tumor Suppressor Proteins ,fibrosis ,Mesenchymal Stem Cells ,medicine.disease ,biology.organism_classification ,Mice, Inbred C57BL ,MicroRNAs ,030104 developmental biology ,Adipose Tissue ,Culture Media, Conditioned ,Reperfusion Injury ,030220 oncology & carcinogenesis ,Reperfusion ,biology.protein ,Apoptosis Regulatory Proteins ,business ,Research Paper - Abstract
Rationale: Cardiovascular diseases, such as myocardial infarction (MI), are the leading causes of death worldwide. Reperfusion therapy is the common standard treatment for MI. However, myocardial ischemia/reperfusion (I/R) causes cardiomyocyte injury, including apoptosis and fibrosis. We aimed to investigate the effects of conditioned medium from adipose-derived stem cells (ADSC-CM) on apoptosis and fibrosis in I/R-treated hearts and hypoxia/reoxygenation (H/R)-treated cardiomyocytes and the underlying mechanisms. Methods: ADSC-CM was collected from ADSCs. The effects of intramuscular injection of ADSC-CM on cardiac function, cardiac apoptosis, and fibrosis examined by echocardiography, Evans blue/TTC staining, TUNEL assay, and Masson's trichrome staining in I/R-treated mice. We also examined the effects of ADSC-CM on apoptosis and fibrosis in H/R-treated H9c2 cells by annexin V/PI flow cytometry, TUNEL assay, and immunocytochemistry. Results: ADSC-CM treatment significantly reduced heart damage and fibrosis of I/R-treated mice and H/R-treated cardiomyocytes. In addition, the expression of apoptosis-related proteins, such as p53 upregulated modulator of apoptosis (PUMA), p-p53 and B-cell lymphoma 2 (BCL2), as well as the fibrosis-related proteins ETS-1, fibronectin and collagen 3, were significantly reduced by ADSC-CM treatment. Moreover, we demonstrated that ADSC-CM contains a large amount of miR-221/222, which can target and regulate PUMA or ETS-1 protein levels. Furthermore, the knockdown of PUMA and ETS-1 decreased the induction of apoptosis and fibrosis, respectively. MiR-221/222 overexpression achieved similar results. We also observed that cardiac I/R markedly increased apoptosis and fibrosis in miR-221/222 knockout (KO) mice, while ADSC-CM decreased these effects. The increased phosphorylation of p38 and NF‐κB not only mediated myocardial apoptosis through the PUMA/p53/BCL2 pathway but also regulated fibrosis through the ETS-1/fibronectin/collagen 3 pathway. Conclusions: Overall, our results show that ADSC-CM attenuates cardiac apoptosis and fibrosis by reducing PUMA and ETS-1 expression, respectively. The protective effect is mediated via the miR-221/222/p38/NF-κB pathway.
- Published
- 2021
15. Holo-lactoferrin: the link between ferroptosis and radiotherapy in triple-negative breast cancer
- Author
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Li-Qiang Qin, Cai-Long Chen, Meng-Lan Lu, Haitao Lv, Kai Yang, Jia-Ying Xu, Zheng Zhang, Xing Tong, and Yun-Hong Li
- Subjects
0301 basic medicine ,China ,Programmed cell death ,DNA damage ,Iron ,Mice, Nude ,Medicine (miscellaneous) ,Breast Neoplasms ,Triple Negative Breast Neoplasms ,SLC7A11 ,Biomarkers, Pharmacological ,Lipid peroxidation ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Holo-Lf ,Cell Line, Tumor ,Tumor Microenvironment ,Animals ,Ferroptosis ,Humans ,skin and connective tissue diseases ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,radiotherapy ,chemistry.chemical_classification ,Mice, Inbred BALB C ,Reactive oxygen species ,Cell Death ,biology ,hypoxia ,Glutathione ,Malondialdehyde ,Lactoferrin ,030104 developmental biology ,chemistry ,Cell culture ,030220 oncology & carcinogenesis ,triple-negative breast cancer ,MCF-7 Cells ,Cancer research ,biology.protein ,Female ,Lipid Peroxidation ,Reactive Oxygen Species ,Research Paper - Abstract
Rationale: Iron-saturated Lf (Holo-Lactoferrin, Holo-Lf) exhibits a superior anticancer property than low iron-saturated Lf (Apo-Lf). Ferroptosis is an iron-dependent cell death characterized by the accumulation of lipid peroxidation products and lethal reactive oxygen species (ROS). Radiotherapy also exerts its therapeutic effect through ROS. Methods: The effect of different iron-saturated Lf on ferroptosis and radiotherapy were tested on triple-negative breast cancer (TNBC) cell line MDA-MB-231 and non-TNBC cell line MCF-7. Results: Holo-Lf significantly increased the total iron content, promoted ROS generation, increased lipid peroxidation end product, malondialdehyde (MDA), and enhanced ferroptosis of MDA-MB-231 cells. By contrast, Apo-Lf upregulated SLC7a11 expression, increased GSH generation and inhibited ferroptosis of MDA-MB-231 cells. However, non-TNBC MCF-7 cells were resistant to Holo-Lf-induced ferroptosis because MCF-7 cells have a higher redox balance capacity than MDA-MB-231 cells. More importantly, Holo-Lf downregulated HIF-1α expression, ameliorated the hypoxia microenvironment in subcutaneous MDA-MB-231 tumors, and promoted radiation-induced DNA damage to hypoxic MDA-MB-231 cells. Finally, the efficacy of radiotherapy to MDA-MB-231 tumors was enhanced by Holo-Lf. Conclusion: Holo-Lf could induce ferroptosis in MDA-MB-231 cells and sensitize MDA-MB-231 tumors to radiotherapy.
- Published
- 2021
16. Compensatory combination of mTOR and TrxR inhibitors to cause oxidative stress and regression of tumors
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Tingting Zhang, Ri Cui, Jundixia Chen, Lin Hong, Fengjiao Wu, Fang Wu, Yiqun Xia, Yun Yu, Chenyu Qiu, Wei Chen, Xin Shen, Chenxin Xu, Congying Xie, Peng Zou, Rongrong Shao, and Peisen Zheng
- Subjects
0301 basic medicine ,autophagy ,Programmed cell death ,Thioredoxin-Disulfide Reductase ,Auranofin ,MAP Kinase Signaling System ,Mice, Nude ,Medicine (miscellaneous) ,Mice ,03 medical and health sciences ,0302 clinical medicine ,c-Jun N-terminal Kinase ,Cell Line, Tumor ,medicine ,Animals ,Humans ,Protein Kinase Inhibitors ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,PI3K/AKT/mTOR pathway ,Mice, Inbred BALB C ,Everolimus ,Cell Death ,business.industry ,TOR Serine-Threonine Kinases ,Autophagy ,Cancer ,thioredoxin reductase ,Endoplasmic Reticulum Stress ,HCT116 Cells ,medicine.disease ,Oxidative Stress ,030104 developmental biology ,030220 oncology & carcinogenesis ,Cancer cell ,mTOR ,Cancer research ,Signal transduction ,business ,Research Paper ,Signal Transduction ,medicine.drug - Abstract
Background: Cancer is a leading cause of death worldwide. Extensive research over decades has led to the development of therapies that inhibit oncogenic signaling pathways. The mammalian target of rapamycin (mTOR) signaling pathway plays an important role in the development of many cancers. Several mTOR inhibitors are approved for the treatment of cancers. However, the anticancer efficacies of mTOR inhibitor monotherapy are still limited. Methods: Western blot was used to detect the expression of indicated molecules. Thioredoxin reductase (TrxR) activity in cells was determined by the endpoint insulin reduction assay. Immunofluorescence staining was used to analyze precise location and expression of target proteins. Nude mice were used for xenograft tumor models. Results: We identified a synergistic lethal interaction of mTOR and TrxR inhibitors and elucidated the underlying molecular mechanisms of this synergism. We demonstrated that mTOR and TrxR inhibitors cooperated to induce cell death by triggering oxidative stress, which led to activation of autophagy, endoplasmic reticulum (ER) stress and c-Jun N-terminal Kinase (JNK) signaling pathway in cancer cells. Remarkably, we found that auranofin (AF) combined with everolimus significantly suppressed tumor growth in HCT116 and SGC-7901 xenograft models with no significant signs of toxicity. Conclusion: Our findings identify a promising therapeutic combination for cancer and has important implications for developing mTOR inhibitor-based combination treatments.
- Published
- 2021
17. Identification of a novel and potent small molecule inhibitor of SRPK1: mechanism of dual inhibition of SRPK1 for the inhibition of cancer progression
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Imteyaz Qamar, Nagendra Singh, Anshuman Chandra, and Hanumappa Ananda
- Subjects
Aging ,flow cytometery ,Cell Survival ,Apoptosis ,Protein Serine-Threonine Kinases ,SRPK1 ,environment and public health ,Serine ,Inhibitory Concentration 50 ,Jurkat Cells ,Humans ,RNA, Messenger ,Phosphorylation ,Binding site ,Protein kinase A ,Protein Kinase Inhibitors ,Cell Proliferation ,chemistry.chemical_classification ,MTT assay ,Cell Death ,Serine-Arginine Splicing Factors ,Kinase ,splicing inhibitor ,Cell Biology ,virtual screening ,Small molecule ,Cell biology ,Molecular Docking Simulation ,Enzyme ,chemistry ,A549 Cells ,structure based drug design ,Drug Design ,Drug Screening Assays, Antitumor ,K562 Cells ,Research Paper ,HeLa Cells - Abstract
Protein kinases are the family of attractive enzyme targets for drug design with relevance to cancer biology. Serine arginine protein kinase 1 (SRPK1) is responsible for the phosphorylation of serine/arginine (SR)-rich proteins. Alternative Splicing Factor/Splicing Factor 2 (ASF/SF2) involved in mRNA editing. ASF/SF2 is over expressed in many cancers and plays crucial roles in the cell survival. Phosphorylation of ASF/SF2 is decisive for its functions in cancer. In search of potential anticancer therapeutic agents for attenuating phosphorylation of ASF/SF2, we have explored specific and potential inhibitors of SRPK1 from natural and drug like compounds databases using in-silico methods. Compound ZINC02154892 (C02) was found to be the most potent inhibitor for SRPK1. In-vitro molecular and cell biology studies have shown C02 as a potent and specific inhibitor of phosphorylation of ASF/SF2 and cell survival in leukemic cell line. Structural analysis of SRPK1 with compound C02 revealed a unique pattern of binding targeting ATP binding site along with inhibiting recruitment of ASF/SF2 by SRPK1. The possibilities of compound C02 to be used as a lead compound paving way for the development of potent and specific inhibitors of SRPK1 for designing of novel potential anticancer inhibitor is inferred from the current studies.
- Published
- 2020
18. An Arabidopsis downy mildew non-RxLR effector suppresses induced plant cell death to promote biotroph infection
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Arne Weiberg, Adriana Trutzenberg, Florian Dunker, Bernhard Lederer, and Lorenz Oberkofler
- Subjects
obligate biotrophic plant parasite ,Arabidopsis thaliana ,host-induced gene silencing (HIGS) ,Physiology ,Arabidopsis ,Hyaloperonospora arabidopsidis ,Nicotiana benthamiana ,Plant Science ,Microbiology ,Gene Expression Regulation, Plant ,Pseudomonas syringae ,Plant Diseases ,Oomycete ,Cell Death ,biology ,AcademicSubjects/SCI01210 ,downy mildew ,Effector ,fungi ,food and beverages ,biology.organism_classification ,non-RxLR cysteine-rich protein effectors (CRs) ,Oomycetes ,Plant—Environment Interactions ,Host-Pathogen Interactions ,Ectopic expression ,Botrytis ,Research Paper - Abstract
Our understanding of obligate biotrophic pathogens is limited by lack of knowledge concerning the molecular function of virulence factors. We established Arabidopsis host-induced gene silencing (HIGS) to explore gene functions of Hyaloperonospora arabidopsidis, including CYSTEINE-RICH PROTEIN (HaCR)1, a potential secreted effector gene of this obligate biotrophic pathogen. HaCR1 HIGS resulted in H. arabidopsidis-induced local plant cell death and reduced pathogen reproduction. We functionally characterized HaCR1 by ectopic expression in Nicotiana benthamiana. HaCR1 was capable of inhibiting effector-triggered plant cell death. Consistent with this, HaCR1 expression in N. benthamiana led to stronger disease symptoms caused by the hemibiotrophic oomycete pathogen Phytophthora capsici, but reduced disease symptoms caused by the necrotrophic fungal pathogen Botrytis cinerea. Expressing HaCR1 in transgenic Arabidopsis confirmed higher susceptibility to H. arabidopsidis and to the bacterial hemibiotrophic pathogen Pseudomonas syringae. Increased H. arabidopsidis infection was in accordance with reduced PATHOGENESIS RELATED (PR)1 induction. Expression of full-length HaCR1 was required for its function, which was lost if the signal peptide was deleted, suggesting its site of action in the plant apoplast. This study provides phytopathological and molecular evidence for the importance of this widespread, but largely unexplored class of non-RxLR effectors in biotrophic oomycetes., Induced cell death is a crucial component of plant defence. An extracellular downy mildew cysteine-rich protein effector inhibited plant cell death enabling host colonization.
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- 2020
19. TNF-α response in macrophages depends on clinical
- Author
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Johann, Guillemot, Christophe, Ginevra, Camille, Allam, Elisabeth, Kay, Christophe, Gilbert, Patricia, Doublet, Sophie, Jarraud, and Annelise, Chapalain
- Subjects
sequence type ,Genotype ,ST1 ,Tumor Necrosis Factor-alpha ,clinical isolates ,Macrophages ,immune response ,Legionella pneumophila ,macrophages ,cell death ,ST47 ,TNF-α ,Humans ,Tumor Necrosis Factor Inhibitors ,Legionnaires' Disease ,Research Article ,Research Paper - Abstract
Legionnaires’ Disease (LD) is a severe pneumonia mainly caused in Europe by Legionella pneumophila serogroup 1 (Lp1). Sequence-based typing methods reveal that some sequence types (ST) are overrepresented in clinical samples such as ST1 and ST47, suggesting that some strains are more fit for infection than others. In the present study, a collection of 108 Lp1 clinical isolates were used to evaluate the strain-dependent immune responses from human macrophages. Clinical Lp1 isolates induced differential TNFα secretion from macrophages. ST1 isolates induced a significantly higher TNF-α secretion than non-ST1, whereas ST47 isolates induced a significantly lower TNF-α secretion than non-ST47 isolates. ST1 isolates induced a significantly higher cell death than ST47 isolates evaluated by lactate dehydrogenase activity (cytotoxicity) and caspase-3 activity (apoptosis). Treatment of macrophages with anti-TNF-α antibodies significantly reduced the cell death in macrophages infected with ST1 or ST47 strains. The TNF-α secretion was neither explained by a differential bacterial replication nor by the number or type (bystander or infected) of TNF-α producing cells following infection but by a differential response from macrophages. The Paris ST1 reference strain elicited a significantly higher TNF-α gene transcription and a higher induction of NF-κB signaling pathway than the Lorraine ST47 reference strain. Clinical Lp1 isolates induce a diverse immune response and cell death, which could be related to the genotype. The two predominant sequence-types ST1 and ST47 trigger opposite inflammatory response that could be related to the host susceptibility.
- Published
- 2022
20. Transient receptor potential ankyrin 1 contributes to the lysophosphatidylcholine-induced oxidative stress and cytotoxicity in OLN-93 oligodendrocyte
- Author
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Hualin Huang, Shuai Li, Feng Tang, Zuoxian Lin, Huifang Zhao, Sihao Deng, Li Zhiyuan, Xiaobo Han, Chao Tian, Lang He, Rongqi Huang, and Xu Junjie
- Subjects
0301 basic medicine ,Nitric Oxide Synthase Type I ,Mitochondrion ,Nitric Oxide ,medicine.disease_cause ,Biochemistry ,Cell Line ,03 medical and health sciences ,chemistry.chemical_compound ,Transient receptor potential channel ,0302 clinical medicine ,Calcium imaging ,Superoxides ,medicine ,Animals ,TRPA1 Cation Channel ,Cell damage ,Membrane Potential, Mitochondrial ,chemistry.chemical_classification ,Original Paper ,Reactive oxygen species ,Cell Death ,Lysophosphatidylcholines ,food and beverages ,Cell Biology ,medicine.disease ,Oligodendrocyte ,Mitochondria ,Rats ,Cell biology ,Oligodendroglia ,Oxidative Stress ,030104 developmental biology ,medicine.anatomical_structure ,Lysophosphatidylcholine ,chemistry ,Calcium ,lipids (amino acids, peptides, and proteins) ,psychological phenomena and processes ,030217 neurology & neurosurgery ,Oxidative stress - Abstract
Transient receptor potential ankyrin 1 (TRPA1), the non-selective cation channel, was found that can mediate the generation of multiple sclerosis, while the mechanism is still controversial. Lysophosphatidylcholine (LPC) is a critical trigger of multiple sclerosis which results from the syndrome of neuronal inflammation and demyelination. In this work, we suggested that TRPA1 can mediate the LPC-induced oxidative stress and cytotoxicity in OLN-93 oligodendrocyte. The expression of TRPA1 in OLN-93 was detected by using quantitative real-time PCR (qRT-PCR) and immunofluorescence. The calcium overload induced by LPC via TRPA1 was detected by calcium imaging. The mechanism of LPC-induced mitochondrial reactive oxygen species (mtROS) generation, mitochondria membrane depolarization, nitric oxide (NO) increase, and development of superoxide production via TRPA1 was verified by using confocal imaging. The cell injury elicited by LPC via TRPA1 was confirmed by both CCK-8 and LDH cytotoxicity detection. These results indicate that TRPA1 plays an important role of the LPC-induced oxidative stress and cell damage in OLN-93 oligodendrocyte. Therefore, inhibition of TRPA1 may protect the LPC-induced demyelination.
- Published
- 2020
21. Autophagy and post-ischemic conditioning in retinal ischemia
- Author
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Leianne Torres, Sophie Tran, Clara R. Stelman, Maryna Salkovski, Figen A. Seiler, Monica Sharma, Mohansrinivas Chennakesavalu, Konrad Kadzielawa, Biji Mathew, Raj Patel, Leslie N. Aldrich, Steven Roth, and Nathan Burg
- Subjects
Male ,0301 basic medicine ,Pathology ,medicine.medical_specialty ,Biology ,03 medical and health sciences ,Autophagy ,medicine ,Animals ,Rats, Wistar ,Ischemic Postconditioning ,Molecular Biology ,PI3K/AKT/mTOR pathway ,Retina ,TUNEL assay ,Cell Death ,030102 biochemistry & molecular biology ,Mechanism (biology) ,Retinal ischemia ,Cell Biology ,Diabetic retinopathy ,medicine.disease ,Oxygen ,Glucose ,030104 developmental biology ,medicine.anatomical_structure ,Astrocytes ,Central retinal artery occlusion ,Beclin-1 ,sense organs ,Lysosomes ,Research Paper - Abstract
Retinal ischemia is a major cause of vision loss and a common underlying mechanism associated with diseases, such as diabetic retinopathy and central retinal artery occlusion. We have previously demonstrated the robust neuroprotection in retina induced by post-conditioning (post-C), a brief period of ischemia, 24 h, following a prolonged and damaging initial ischemia. The mechanisms underlying post-C-mediated retinal protection are largely uncharacterized. We hypothesized that macroautophagy/autophagy is a mediator of post-C-induced neuroprotection. This study employed an in vitro model of oxygen glucose deprivation (OGD) in the retinal R28 neuronal cell line, and an in vivo rat model of retinal ischemic injury. In vivo, there were significant increases in autophagy proteins, MAP1LC3-II/LC3-II, and decreases in SQSTM1/p62 (sequestosome 1) in ischemia/post-C vs. ischemia/sham post-C. Blockade of Atg5 and Atg7 in vivo decreased LC3-II, increased SQSTM1, attenuated the functional protective effect of post-C, and increased histological damage and TUNEL compared to non-silencing siRNA. TUNEL after ischemia in vivo was found in retinal ganglion, amacrine, and photoreceptor cells. Blockade of Atg5 attenuated the post-C neuroprotection by a brief period of OGD in vitro. Moreover, in vitro, post-C attenuated cell death, loss of cellular proliferation, and defective autophagic flux from prolonged OGD. Stimulating autophagy using Tat-Beclin 1 rescued retinal neurons from cell death after OGD. As a whole, our results suggest that autophagy is required for the neuroprotective effect of retinal ischemic post-conditioning and augmentation of autophagy offers promise in the treatment of retinal ischemic injury. Abbreviations: BECN1: Beclin 1, autophagy related; DAPI: 4’,6-diamidino-2-phenylindole; DR: diabetic retinopathy; EdU: 5-ethynyl-2’-deoxyuridine; ERG: Electroretinogram; FITC: Fluorescein isothiocyanate; GCL: Ganglion cell layer; GFAP: Glial fibrillary acidic protein; INL: Inner nuclear layer; IPL: Inner plexiform layer; MAP1LC3/LC3: Microtubule-associated protein 1 light chain 3; OGD: Oxygen-glucose deprivation; ONL: Outer nuclear layer; OP: Oscillatory potential; PFA: Paraformaldehyde; PL: Photoreceptor layer; post-C: post-conditioning; RFP: Red fluorescent protein; RGC: Retinal ganglion cell; RPE: Retinal pigment epithelium; RT-PCR: Real-time polymerase chain reaction; SEM: Standard error of the mean; siRNA: Small interfering RNA; SQSTM1: Sequestosome 1; STR: Scotopic threshold response; Tat: Trans-activator of transcription; TUNEL: Terminal deoxynucleotidyl transferase dUTP nick end labeling
- Published
- 2020
22. Stress-induced changes in CARF expression determine cell fate to death, survival, or malignant transformation
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Caroline T. Cheung, Rajkumar Singh Kalra, Amr Omar, Sunil C. Kaul, Sukant Garg, Renu Wadhwa, and Anupama Chaudhary
- Subjects
0301 basic medicine ,Senescence ,Programmed cell death ,Cell Survival ,Cell ,Cell fate determination ,Biology ,Biochemistry ,Cell Line ,Malignant transformation ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Stress, Physiological ,medicine ,Animals ,Humans ,Cell Proliferation ,Original Paper ,Cell Death ,RNA-Binding Proteins ,Cell Biology ,In vitro ,Telomere ,Cell biology ,Cell Transformation, Neoplastic ,030104 developmental biology ,medicine.anatomical_structure ,Apoptosis ,030220 oncology & carcinogenesis ,NIH 3T3 Cells ,Apoptosis Regulatory Proteins ,Biomarkers - Abstract
CARF (Collaborator of ARF) was discovered as an ARF-interacting protein that activated ARF-p53-p21(WAF1) signaling involved in cellular response to a variety of stresses, including oxidative, genotoxic, oncogenic, or telomere deprotection stresses, leading to senescence, growth arrest, or apoptosis. Of note, whereas suppression of CARF was lethal, its enrichment was associated with increased proliferation and malignant transformation of cells. These reports have predicted that CARF could serve as a multi-stress marker with a predictive value for cell fates. Here, we recruited various in vitro stress models and examined their effect on CARF expression using human normal fibroblasts. We demonstrate that CARF levels in stress and post-stress conditions could predict the fate of cells towards either death or enhanced proliferation and malignant transformation. We provide extensive molecular evidence that (i) CARF expression changes in response to stress, (ii) it modulates cell death or survival signaling and determines the fate of cells, and (iii) it may serve as a predictive measure of cellular response to stress and an important marker for biosafety. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s12192-020-01088-y) contains supplementary material, which is available to authorized users.
- Published
- 2020
23. Testosterone increases apoptotic cell death and decreases mitophagy in Leber’s hereditary optic neuropathy cells
- Author
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Parvana Hajieva, Magdalena Korwin, Elona Jankauskaitė, Katarzyna Tońska, Anna M. Ambroziak, Monika Ołdak, Ewa Bartnik, and Agata Kodroń
- Subjects
Cell death ,Adult ,Male ,Retinal Ganglion Cells ,0301 basic medicine ,Mitochondrial DNA ,Programmed cell death ,Apoptosis ,Optic Atrophy, Hereditary, Leber ,Biology ,medicine.disease_cause ,DNA, Mitochondrial ,Retinal ganglion ,LHON ,03 medical and health sciences ,Human Genetics • Original Paper ,0302 clinical medicine ,Mitophagy ,Autophagy ,Genetics ,medicine ,Humans ,Point Mutation ,Testosterone ,Mutation ,Blood Cells ,Leber's hereditary optic neuropathy ,General Medicine ,medicine.disease ,Mitochondria ,Nucleosomes ,030104 developmental biology ,Genome, Mitochondrial ,Cancer research ,Female ,Macrolides ,030217 neurology & neurosurgery - Abstract
Leber’s hereditary optic neuropathy (LHON) is one of the most common mitochondrial diseases caused by point mutations in mitochondrial DNA (mtDNA). The majority of diagnosed LHON cases are caused by a point mutation at position 11,778 in the mitochondrial genome. LHON mainly affects young men in their 20s and 30s with usually poor visual prognosis. It remains unexplained why men are more likely to develop the disease and why only retinal ganglion cells are affected. In this study, a cell model was used for the first time to investigate the influence of testosterone on the cell death mechanism apoptosis and on an autophagy/mitophagy. Cells with m.11778G > A were found to be significantly more susceptible to nucleosome formation and effector caspase activation that serve as hallmarks of apoptotic cell death. Cells having this mutation expressed higher levels of mitophagic receptors BNIP3 and BNIP3L/Nix in a medium with testosterone. Moreover, cells having the mutation exhibited greater mitochondrial mass, which suggests these cells have a decreased cell survival. The observed decrease in cell survival was supported by the observed increase in apoptotic cell death. Autophagy was analyzed after inhibition with Bafilomycin A1 (Baf A1). The results indicate impairment in autophagy in LHON cells due to lower autophagic flux supported by observed lower levels of autophagosome marker LC3-II. The observed impaired lower autophagic flux in mutant cells correlated with increased levels of BNIP3 and BNIP3L/Nix in mutant cells.
- Published
- 2020
24. Berberine attenuates mitochondrial dysfunction by inducing autophagic flux in myocardial hypoxia/reoxygenation injury
- Author
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Xueming Cao, Jiang Li, Jing Zhang, Yuwei Zhang, Peiyuan Hao, Li Li, Yan Chen, Na Zhu, and Chuanyu Gao
- Subjects
0301 basic medicine ,Berberine ,Apoptosis ,Myocardial Reperfusion Injury ,Pharmacology ,Biochemistry ,Flow cytometry ,Green fluorescent protein ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Downregulation and upregulation ,Autophagy ,medicine ,Animals ,Myocytes, Cardiac ,Cells, Cultured ,Membrane Potential, Mitochondrial ,Membrane potential ,Original Paper ,Cell Death ,medicine.diagnostic_test ,Cell Biology ,Hypoxia (medical) ,Fusion protein ,Mitochondria ,Rats ,030104 developmental biology ,chemistry ,030220 oncology & carcinogenesis ,medicine.symptom - Abstract
Berberine (BBR) is routinely prescribed in many Asian countries to treat diarrhea. Evidence from both animal and clinical investigations suggests that BBR exerts diverse pharmacological activities, including antidiabetic, antineoplastic, antihypertensive, and antiatherosclerotic effects. This study aimed to explore the cardioprotective mechanisms of BBR and to elucidate the modulations between autophagy and mitochondrial function during hypoxia/reoxygenation (H/R) in H9c2 cells. The degree of autophagic flux was assessed by pretreating H9c2 cells with BBR prior to H/R exposure and measuring the expression levels of Beclin-1 and green fluorescent protein (GFP)-labeled LC3B fusion proteins as well as the LC3II/LC3I ratio. The mitochondrial membrane potential (△Ψm) in H9c2 cells was evaluated by detecting rhodamine-123 fluorescence using flow cytometry. The results revealed that pretreatment with BBR upregulated autophagic flux and protected against the loss of the △Ψm in H9c2 cells subjected to H/R. We conclude that BBR attenuates mitochondrial dysfunction by inducing autophagic flux.
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- 2020
25. Patient-derived glioblastoma cultures as a tool for small-molecule drug discovery
- Author
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Kimberly Malesky, Donald C. Lo, Guizhi Yang, Alice Loo, Hui Gao, Joshua M. Korn, Raymond Pagliarini, Brent R. Stockwell, Fallon Lin, Tom Mikkelsen, Eduard Reznik, Ana C. deCarvalho, and Ling F. Ye
- Subjects
0301 basic medicine ,medicine.medical_treatment ,chemical biology ,Biology ,Targeted therapy ,03 medical and health sciences ,0302 clinical medicine ,glioma ,Neurosphere ,Glioma ,Gene expression ,medicine ,cancer ,Drug discovery ,RNA ,Cancer ,medicine.disease ,nervous system diseases ,3. Good health ,cell death ,030104 developmental biology ,Oncology ,Cell culture ,030220 oncology & carcinogenesis ,Cancer research ,Research Paper - Abstract
There is a compelling need for new therapeutic strategies for glioblastoma multiforme (GBM). Preclinical target and therapeutic discovery for GBMs is primarily conducted using cell lines grown in serum-containing media, such as U-87 MG, which do not reflect the gene expression profiles of tumors found in GBM patients. To address this lack of representative models, we sought to develop a panel of patient-derived GBM models and characterize their genomic features, using RNA sequencing (RNA-seq) and growth characteristics, both when grown as neurospheres in culture, and grown orthotopically as xenografts in mice. When we compared these with commonly used GBM cell lines in the Cancer Cell Line Encyclopedia (CCLE), we found these patient-derived models to have greater diversity in gene expression and to better correspond to GBMs directly sequenced from patient tumor samples. We also evaluated the potential of these models for targeted therapy, by using the genomic characterization to identify small molecules that inhibit the growth of distinct subsets of GBMs, paving the way for precision medicines for GBM.
- Published
- 2020
26. Mouse skin-derived precursors alleviates ultraviolet B irradiation damage via early activation of TGF-β/Smad pathway by thrombospondin1
- Author
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Ru Dai, Lidan Xiong, Jie Tang, Guonian Zhu, Li Li, and Yiming Li
- Subjects
0301 basic medicine ,Cell Survival ,Ultraviolet Rays ,Cell ,Smad Proteins ,SMAD ,Biology ,Models, Biological ,Collagen Type I ,Thrombospondin 1 ,Extracellular matrix ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Transforming Growth Factor beta ,medicine ,Animals ,Viability assay ,Cell Shape ,Molecular Biology ,Cellular Senescence ,Skin ,Cell Death ,integumentary system ,Stem Cells ,Mesenchymal stem cell ,Cell Biology ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Matrix Metalloproteinase 1 ,Signal transduction ,Research Paper ,Signal Transduction ,Developmental Biology ,Transforming growth factor - Abstract
Our previous research implied mouse skin-derived precursors (mSKPs) possessed the capacity of anti-ultraviolet B (UVB) irradiation damage, and the mechanisms might be associated with transforming growth factor-β (TGF-β) signaling pathway activation. In this study, we investigated and compared the response to UVB irradiation between mSKPs and dermal mesenchymal stem cells (dMSCs), and explored the underlying mechanisms. Irradiation damage such as decreased cell viability, cell senescence, and cell death was observed in both mSKPs and dMSCs at 24 h after UVB exposure. In mSKPs, change in cell morphology, viability, cell senescence and death at the following time points implied the recovery of UVB irradiation damage. Additionally, thrombospondin1 (TSP1) and TGF-β1 increased significantly in mSKPs’ supernatant after UVB irradiation. The gene expression of TSP1, TGF-β1, metalloproteinase 1 (MMP1), and Collagen I elevated shortly after the UVB exposure. The protein expression of TSP1, TGF-β1, MMP1, Collagen I, smad2/3, and p-smad2/3 at multiple time points after the UVB exposure was consistent with the gene expression results. In dMSCs, no obvious recovery was noticed. Together, these results revealed that in mSKPs, one of the mechanisms to attenuate the UVB irradiation damage might be the early activation of TGF-β/Smad pathway by TSP1. Given that mSKPs could differentiate into fibroblast-like SKP-derived fibroblasts (SFBs) in vivo or with the presence of serum, mSKPs might serve as a therapeutic potential for fibroblasts supplement and UVB irradiation damage treatment. Abbreviations: SKPs: skin-derived precursors; mSKPs: mouse SKPs; UVB: ultraviolet B; TGF-β/Smad: transforming growth factor-β/Smad; TSP1: thrombospondin 1; MMP 13: metalloproteinases 13; TβRII: TGF-β receptor II; SFBs: SKP-derived fibroblasts; KEGG: Kyoto encyclopedia of genes and genomes; DEGs: differentially expressed genes; dMSCs: dermal mesenchymal stem cells; LM: light microscope; CCK-8: cell counting kit 8; ELISA: Enzyme-linked immuno sorbent assay; qRT-PCR: quantitative real-time polymerase chain reaction; TSPs: thrombospondins; ECM: extracellular matrix; R-smads: receptor-regulated smads
- Published
- 2020
27. Fibroblast growth factor 10 alleviates particulate matter-induced lung injury by inhibiting the HMGB1-TLR4 pathway
- Author
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Junjie Chen, Yiran Hu, Chen Chaolei, Lingjing Liu, Jin-San Zhang, Mingyang Zhu, Chenjian Song, Nian Dong, Chengshui Chen, Qiang Wang, and Jingli Li
- Subjects
Aging ,Biopsy ,Gene Expression ,Inflammation ,Respiratory Mucosa ,Lung injury ,Pharmacology ,HMGB1 ,medicine ,Humans ,TLR4 ,HMGB1 Protein ,particulate matter ,FGF10 ,Lung ,Cell Death ,medicine.diagnostic_test ,biology ,Chemistry ,Interleukin ,Lung Injury ,Cell Biology ,Immunohistochemistry ,Toll-Like Receptor 4 ,stomatognathic diseases ,Bronchoalveolar lavage ,medicine.anatomical_structure ,inflammation ,Gene Knockdown Techniques ,biology.protein ,Cytokines ,Tumor necrosis factor alpha ,Disease Susceptibility ,medicine.symptom ,Fibroblast Growth Factor 10 ,Biomarkers ,Research Paper ,Signal Transduction - Abstract
Exposure to particulate matter (PM) is associated with increased incidence of respiratory diseases. The present study aimed to investigate the roles of fibroblast growth factor 10 (FGF10) in PM-induced lung injury. Mice were intratracheally instilled with FGF10 or phosphate-buffered saline at one hour before instillation of PM for two consecutive days. In addition, the anti-inflammatory impact of FGF10 in vitro and its effect on the high-mobility group box 1 (HMGB1)-toll-like receptor 4 (TLR4) pathway was investigated. It was found that PM exposure is associated with increased inflammatory cell infiltration into the lung and increased vascular protein leakage, while FGF10 pretreatment attenuated both of these effects. FGF10 also decreased the PM-induced expression of interleukin (IL)-6, IL-8, tumor necrosis factor-α and HMGB1 in murine bronchoalveolar lavage fluid and in the supernatants of human bronchial epithelial cells exposed to PM. FGF10 exerted anti-inflammatory and cytoprotective effects by inhibiting the HMGB1-TLR4 pathway. These results indicate that FGF10 may have therapeutic values for PM-induced lung injury.
- Published
- 2020
28. Circular RNA HIPK3 downregulation mediates hydrogen peroxide-induced cytotoxicity in human osteoblasts
- Author
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Chong Chen, Hong Zhao, Xiang-yang Zhang, Jinqian Liang, Jianhua Hu, and Ying-Chao Shen
- Subjects
musculoskeletal diseases ,Aging ,Programmed cell death ,oxidative injury ,Cell Survival ,hydrogen peroxide ,Apoptosis ,Protein Serine-Threonine Kinases ,Ectopic Gene Expression ,Small hairpin RNA ,Downregulation and upregulation ,Humans ,RNA, Small Interfering ,Cytotoxicity ,Protein kinase A ,Cells, Cultured ,Osteoblasts ,Cell Death ,biology ,Chemistry ,Intracellular Signaling Peptides and Proteins ,RNA, Circular ,Cell Biology ,Transfection ,circular RNA HIPK3 ,Cell biology ,Gene Expression Regulation ,biology.protein ,microRNA-124 ,RNA Interference ,Cyclin-dependent kinase 6 ,Research Paper - Abstract
Hydrogen peroxide (H2O2) induces oxidative injury to human osteoblasts. The expression and potential function of circular RNA HIPK3 (circHIPK3) in H2O2-treated human osteoblasts were tested. We show that H2O2 significantly downregulated circHIPK3 in OB-6 cells and primary human osteoblasts. Furthermore, circHIPK3 levels were decreased in the necrotic femoral head tissues of dexamethasone-treated patients. In OB-6 osteoblastic cells and primary human osteoblasts, forced overexpression of circHIPK3 by a lentiviral construct alleviated H2O2-induced viability reduction, cell death and apoptosis. Contrarily, circHIPK3 silencing by targeted shRNA potentiated H2O2-induced cytotoxicity in OB-6 cells and primary human osteoblasts. Moreover, circHIPK3 downregulation by H2O2 induced miR-124 accumulation in OB-6 cells and primary human osteoblasts. On the contrary, miR-124 inhibition by transfection of the miR-124 inhibitor protected human osteoblasts from H2O2. Importantly, forced overexpression of miR-124 by transfection of the miR-124 mimic induced significant cytotoxicity in OB-6 cells and primary human osteoblasts. H2O2 downregulated miR-124’s targets, cyclin dependent kinase 6 and Rho-Associated Protein Kinase 1, in human osteoblasts. In conclusion circHIPK3 downregulation mediates H2O2-induced cytotoxicity in human osteoblasts.
- Published
- 2020
29. Histone deacetylase inhibitor MPT0B291 suppresses Glioma Growth in vitro and in vivo partially through acetylation of p53
- Author
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Batsaikhan Buyandelger, Huei Mei Huang, Ruei Ming Chen, Kuo Sheng Hung, Jing Ping Liou, Yung Hsiao Chiang, Eli E. Bar, and Jia Yi Wang
- Subjects
Male ,allograft ,Programmed cell death ,Cell cycle checkpoint ,medicine.drug_class ,Mice, Nude ,Antineoplastic Agents ,Applied Microbiology and Biotechnology ,Rats, Sprague-Dawley ,Mice ,03 medical and health sciences ,Cell Line, Tumor ,Glioma ,medicine ,Animals ,Humans ,xenograft ,Phosphorylation ,Molecular Biology ,Ecology, Evolution, Behavior and Systematics ,030304 developmental biology ,0303 health sciences ,Cell Death ,Chemistry ,HDAC6 inhibition ,Histone deacetylase inhibitor ,Acetylation ,Cell Biology ,Cell cycle ,medicine.disease ,G1 Phase Cell Cycle Checkpoints ,Xenograft Model Antitumor Assays ,Rats ,Histone Deacetylase Inhibitors ,cell cycle arrest ,Apoptosis ,Astrocytes ,Cancer research ,Histone deacetylase ,Tumor Suppressor Protein p53 ,Research Paper ,Developmental Biology - Abstract
Background: Histone deacetylase (HDAC) inhibitors have emerged as a new class of anti-tumor agents for various types of tumors, including glioblastoma. Methods and results: We found that a novel HDAC inhibitor, MPT0B291, significantly reduced the cell viability and increased cell death of human and rat glioma cell lines, but not in normal astrocytes. We also demonstrated that MPT0B291 suppressed proliferation by inducing G1 phase cell cycle arrest and increased apoptosis in human and rat glioma cell lines by flow cytometry and immunocytochemistry. We further investigated the anti-tumor effects of MPT0B291 in xenograft (mouse) and allograft (rat) models. The IVIS200 images and histological analysis indicated MPT0B291 (25 mg/kg, p. o.) reduced tumor volume. Mechanistically, MPT0B291 increased phosphorylation and acetylation/activation of p53 and increased mRNA levels of the apoptosis related genes PUMA, Bax, and Apaf1 as well as increased protein level of PUMA, Apaf1 in C6 cell line. The expression of cell cycle related gene p21 was also increased and Cdk2, Cdk4 were decreased by MPT0B291. Conclusion: Our study highlights the anti-tumor efficacy of a novel compound MPT0B291 on glioma growth.
- Published
- 2020
30. Zyflamend, a unique herbal blend, induces cell death and inhibits adipogenesis through the coordinated regulation of PKA and JNK
- Author
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Dallas Donohoe, Mohammed AlQuraishi, Samah Chahed, Jay Whelan, Dina Alani, Dexter L. Puckett, Victoria D. Frankel, Brynn H. Voy, and Ahmed Bettaieb
- Subjects
Programmed cell death ,2019-20 coronavirus outbreak ,Histology ,Coronavirus disease 2019 (COVID-19) ,Physiology ,Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ,Adipose tissue ,Diseases of the endocrine glands. Clinical endocrinology ,adipogenesis ,ampk ,Mice ,3T3-L1 Cells ,Adipocytes ,Animals ,QP1-981 ,Lipolysis ,Medicine ,jnk ,Cells, Cultured ,Cell Death ,Dose-Response Relationship, Drug ,QH573-671 ,Plant Extracts ,business.industry ,digestive, oral, and skin physiology ,JNK Mitogen-Activated Protein Kinases ,food and beverages ,AMPK ,Cell Differentiation ,Cell Cycle Checkpoints ,Cell Biology ,RC648-665 ,Cyclic AMP-Dependent Protein Kinases ,zyflamend ,Adipogenesis ,lipolysis ,Cancer research ,pka ,Cytology ,business ,Research Article ,Research Paper ,Signal Transduction - Abstract
The prevalence of obesity and its comorbidities has sparked a worldwide concern to address rates of adipose tissue accrual. Recent studies have demonstrated a novel role of Zyflamend, a blend of natural herbal extracts, in regulating lipid metabolism in several cancer cell lines through the activation of the AMPK signalling pathway. Yet, the role of Zyflamend in adipogenic differentiation and lipid metabolism remains largely unexplored. The objective of this study is to investigate the effects of Zyflamend on white 3T3-MBX pre-adipocyte differentiation and elucidate the molecular mechanisms. We demonstrate that Zyflamend treatment altered cell cycle progression, attenuated proliferation, and increased cell death of 3T3-MBX pre-adipocytes. In addition, treatment with Zyflamend inhibited lipid accumulation during the differentiation of 3T3-MBX cells, consistent with decreased expression of lipogenic genes and increased lipolysis. Mechanistically, Zyflamend-induced alterations in adipogenesis were mediated, at least in part, through the activation of AMPK, PKA, and JNK. Inhibition of AMPK partially reversed Zyflamend-induced inhibition of differentiation, whereas the inhibition of either JNK or PKA fully restored adipocyte differentiation and decreased lipolysis. Taken together, the present study demonstrates that Zyflamend, as a novel anti-adipogenic bioactive mix, inhibits adipocyte differentiation through the activation of the PKA and JNK pathways. Abbreviation 7-AAD: 7-amino-actinomycin D; ACC: acetyl-CoA carboxylase; AKT: protein kinase B; AMPK: AMP-activated protein kinase; ATGL: adipose triglyceride lipase; C/EBPα: CCAAT-enhancer binding protein alpha; DMEM: Dulbecco’s Modified Eagle Medium; DMSO: dimethyl sulphoxide; DTT: dithiothreitol; EGTA: ethylene glycol-bis-(2-aminoethyl)-N,N,N’,N’-tetraacetic acid; ERK: extracellular signal–regulated kinases; FASN: fatty acid synthase; FBS: foetal bovine serum; GLUT: glucose transporter; HSL: hormone-sensitive lipase; IR: insulin receptor; IRS: insulin receptor substrate; JNK: c-JUN N-terminal kinase; MGL: monoacylglycerol lipase; NaF: sodium fluoride; NF-κB: nuclear factor kappa-light-chain-enhancer of activated B cells; PBS: phosphate buffered- saline; PCB: pyruvate carboxylase; PDE: phosphodiesterase; PKA: protein kinase cAMP-dependent; PMSF: phenylmethylsulfonyl fluoride; PPARγ: perilipin peroxisome proliferator-activated receptor gamma; PREF-1: pre-adipocyte factor 1; PVDF: polyvinylidene fluoride; RIPA: radio-immunoprecipitation assay; SDS-PAGE: sodium dodecyl sulphate polyacrylamide gel electrophoresis; SEM: standard error of the mean; SOX9: suppressor of cytokine signalling 9; TGs: triacylglycerols.
- Published
- 2020
31. Protective Effects of Foam Rolling against Inflammation and Notexin Induced Muscle Damage in Rats
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Pilar Rivera, Laura Elvira, Francisco Miguel Martínez-Arnau, Carlos Colmena, Soraya L. Valles, Diego Ceca, Adrian Jorda, and Ana Pablos
- Subjects
Muscle tissue ,medicine.medical_specialty ,N-group (finite group theory) ,Inflammation ,Hindlimb ,Muscle damage ,03 medical and health sciences ,Grip strength ,muscle recovery ,0302 clinical medicine ,Physical Conditioning, Animal ,Internal medicine ,foam rolling ,medicine ,Animals ,Humans ,Muscle Strength ,Range of Motion, Articular ,Rats, Wistar ,Muscle, Skeletal ,Physical Therapy Modalities ,Balance (ability) ,Elapid Venoms ,Massage ,Tumor Necrosis Factor-alpha ,Chemistry ,General Medicine ,Rats ,Physical Therapists ,Disease Models, Animal ,cell death ,medicine.anatomical_structure ,Endocrinology ,030211 gastroenterology & hepatology ,Foam rolling ,medicine.symptom ,performance ,Interleukin-1 ,Sports ,Research Paper - Abstract
It is known that high-intensity exercise can cause inflammation and damage in muscle tissue, and in recent years, physical therapists and fitness professionals have begun to use foam rolling as a recovery method to improve performance. Despite the lack of basic science studies to support or refute the efficacy of foam rolling, the technique is very widely used in the sports world. In this respect, we investigated whether foam rolling could attenuate muscle damage and inflammation. Female Wistar rats were assigned to control (C), foam rolling (FR), notexin without foam rolling (N) and notexin with foam rolling (NFR) groups. A 4.5 x 2 cm foam roller was used to massage their hind legs (two 60-second repetitions twice a day for 3 days). Motor function tests (Balance Beam Test and Grip strength) were used. We detected an increase in time and foot faults when crossing a beam in the N group compared to C and FR rats. In contrast, a significant decrease was detected in both tests in NFR compared to N rats. Muscle power was measured with a grip strength test and better performance was detected in NFR rats compared to N rats. Furthermore, an increase of pro-inflammatory proteins was noted in the N group, while there was a decrease in the NFR group. On the contrary, an increase in PPAR-γ (anti-inflammatory protein) in the NFR group compared to the N group demonstrates the anti-inflammatory properties of the foam rolling technique. In summary, applying foam rolling after damage has benefits such as an increase in anti-inflammatory proteins and a reduction of pro-inflammatory proteins, resulting in muscle recovery and better performance.
- Published
- 2020
32. TFE3, a potential therapeutic target for Spinal Cord Injury via augmenting autophagy flux and alleviating ER stress
- Author
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Huanwen Chen, Yuqin Mao, Jing Zhang, Ling Xie, Zhilong Zheng, Mi Liu, Wanying Zhang, Hongyu Zhang, Huazi Xu, Jian Xiao, Wen Han, Kailiang Zhou, and Yao Li
- Subjects
Medicine (miscellaneous) ,Apoptosis ,AMPK signaling pathways ,Spinal cord injury ,02 engineering and technology ,medicine.disease_cause ,PC12 Cells ,Mice ,03 medical and health sciences ,Cell Line, Tumor ,Autophagy ,medicine ,Animals ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,Spinal Cord Injuries ,030304 developmental biology ,Mice, Knockout ,Neurons ,chemistry.chemical_classification ,0303 health sciences ,Reactive oxygen species ,TUNEL assay ,Cell Death ,Basic Helix-Loop-Helix Leucine Zipper Transcription Factors ,TOR Serine-Threonine Kinases ,AMPK ,Recovery of Function ,Endoplasmic Reticulum Stress ,021001 nanoscience & nanotechnology ,ER stress-induced apoptosis ,Rats ,Cell biology ,Mice, Inbred C57BL ,TFE3 ,chemistry ,Unfolded protein response ,Female ,Signal transduction ,Lysosomes ,Reactive Oxygen Species ,0210 nano-technology ,Flux (metabolism) ,Oxidative stress ,Signal Transduction ,Transcription Factors ,Research Paper - Abstract
Background and Aim: Increasing evidence suggests that spinal cord injury (SCI)-induced defects in autophagic flux may contribute to an impaired ability for neurological repair following injury. Transcription factor E3 (TFE3) plays a crucial role in oxidative metabolism, lysosomal homeostasis, and autophagy induction. Here, we investigated the role of TFE3 in modulating autophagy following SCI and explored its impact on neurological recovery. Methods: Histological analysis via HE, Nissl and Mason staining, survival rate analysis, and behavioral testing via BMS and footprint analysis were used to determine functional recovery after SCI. Quantitative real-time polymerase chain reaction, Western blotting, immunofluorescence, TUNEL staining, enzyme-linked immunosorbent assays, and immunoprecipitation were applied to examine levels of autophagy flux, ER-stress-induced apoptosis, oxidative stress, and AMPK related signaling pathways. In vitro studies using PC12 cells were performed to discern the relationship between ROS accumulation and autophagy flux blockade. Results: Our results showed that in SCI, defects in autophagy flux contributes to ER stress, leading to neuronal death. Furthermore, SCI enhances the production of reactive oxygen species (ROS) that induce lysosomal dysfunction to impair autophagy flux. We also showed that TFE3 levels are inversely correlated with ROS levels, and increased TFE3 levels can lead to improved outcomes. Finally, we showed that activation of TFE3 after SCI is partly regulated by AMPK-mTOR and AMPK-SKP2-CARM1 signaling pathways. Conclusions: TFE3 is an important regulator in ROS-mediated autophagy dysfunction following SCI, and TFE3 may serve as a promising target for developing treatments for SCI.
- Published
- 2020
33. Effects of neoadjuvant FOLFIRINOX and gemcitabine-based chemotherapy on cancer cell survival and death in patients with pancreatic ductal adenocarcinoma
- Author
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Milena Sachsenmaier, Markus W. Büchler, Ulla Klaiber, Frank Bergmann, John P. Neoptolemos, Thilo Hackert, Oliver Strobel, Franco Fortunato, Li Xie, Ulf Hinz, and Leizhou Xia
- Subjects
0301 basic medicine ,autophagy ,Programmed cell death ,endocrine system diseases ,FOLFIRINOX ,medicine.medical_treatment ,Necroptosis ,03 medical and health sciences ,0302 clinical medicine ,stroma ,medicine ,neoadjuvant therapy ,Neoadjuvant therapy ,Chemotherapy ,business.industry ,Cancer ,medicine.disease ,Gemcitabine ,cell death ,030104 developmental biology ,Oncology ,030220 oncology & carcinogenesis ,Cancer cell ,Cancer research ,prognosis ,business ,Research Paper ,medicine.drug - Abstract
Background: The progression and response to systemic treatment of cancer is substantially dependent on the balance between cancer cell death (apoptosis and necroptosis) and cancer cell survival (autophagy). Although well characterized in experimental systems, the status of cancer cell survival and cell death in human pancreatic ductal adenocarcinoma (PDAC), especially in response to chemotherapy and different types of chemotherapy is poorly described. Results: The median (95% confidence interval) survival was 31.6 (24.5–44.5) months after FOLFIRINOX versus 15.8 (2.0–20.5) months after gemcitabine-based therapy (p = 0.039). PDAC tissue autophagy was reduced compared to normal pancreata based on reduced BECLIN-1 expression and LC3-Lamp-2 colocalization, whilst necroptosis (RIP-1) was increased. Neoadjuvant therapy was associated with further reduced autophagy based on p62/SQSTM-1 accumulation, and increased necroptosis (RIP3 and pMLKL) and apoptosis (BAX, cleaved CASPASE-9 and CASPASE-3) markers, increased nuclear p65 (NF-κB) and extracellular HMGB1 expression, with greater CD8+ lymphocyte infiltration. Survival was associated with reduced autophagy and increased apoptosis. Necroptosis (RIP-3, pMLKL) and apoptosis (BAX and cleaved CASPASE-9) markers were higher after FOLFIRINOX than gemcitabine-based treatment. Patients and methods: Cancer cell autophagy, apoptosis, and necroptosis marker expression was compared in pancreatic tissue samples from 51 subjects, comprising four groups: (1) surgical resection for PDAC after FOLFIRINOX (n = 11), or (2) after gemcitabine-based (n = 14) neoadjuvant therapy, (3) patients undergoing PDAC resection without prior chemotherapy (n = 13), and (4) normal pancreata from 13 organ donors. Marker expression was undertaken using semi-automated immunofluorescence-FACS-like analysis, defining PDAC cells by CK-7+ expression.
- Published
- 2019
34. Cell membrane breakage and triggering T cell infiltration are involved in human telomerase reverse transcriptase (hTERT) promoter-driven novel peptide KK-64 for liver cancer gene therapy
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Yuanhua Lu, Jie Ma, Jian Lin, Yafei Tian, Yongjun Ma, Wei Wang, Jialin Li, Hugang Zhang, and Ping Jiao
- Subjects
Male ,T-Lymphocytes ,Bioengineering ,Apoptosis ,Applied Microbiology and Biotechnology ,Protein Structure, Secondary ,liver cancer ,KK-64 ,Cell Movement ,Cell Line, Tumor ,Animals ,Humans ,Neoplasm Invasiveness ,Promoter Regions, Genetic ,Telomerase ,Cell Proliferation ,Mice, Inbred BALB C ,ACP ,Cell Death ,Cell Membrane ,Liver Neoplasms ,General Medicine ,Genetic Therapy ,tumor growth ,Peptides ,TP248.13-248.65 ,Biotechnology ,Plasmids ,Research Article ,Research Paper - Abstract
Liver cancer is an aggressive malignancy with exhibits both high mortality and morbidity. The current treatment options are associated with several limitations, novel specific anti-cancer drugs are urgently needed to improve liver cancer treatment. In this study, a new peptide KK-64 was designed, and it showed strong cytotoxicity against liver cancer cells. To obtain the tumor targeting property, a plasmid that contains KK-64 DNA fragment and driven by human telomerase reverse transcriptase (hTERT) promoter was constructed. pcTERT-kk-64 plasmid was found to specifically inhibit the viability of liver cancer cells HepG2, induce substantial apoptosis as well as damage to the cell membranes, but had minimal effects toward normal liver HL-7702 cells. Furthermore, pcTERT-kk-64 plasmids was also noted to significantly attenuate migration and invasion of HepG2 cells. The anti-tumor effect of pcTERT-kk-64 plasmid was also observed in H22 cell-bearing mice, and it appeared to cause significant tumor regression, trigger tumor cell apoptosis, and infiltrate cytotoxicity T cells to the tumor tissues after plasmids injection. Thus, pcTERT-kk-64 plasmids showed both strong cytotoxicity and tumor selectivity in vitro and in tumor-bearing mice in liver cancer models.
- Published
- 2021
35. LAMP3 inhibits autophagy and contributes to cell death by lysosomal membrane permeabilization
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Toshio Odani, Hongen Yin, John A. Chiorini, Hiroyuki Nakamura, Drew G. Michael, Tsutomu Tanaka, Blake M. Warner, Masayuki Noguchi, and Tatsuya Atsumi
- Subjects
Cathepsin ,Programmed cell death ,Cell Membrane Permeability ,LAMP1 ,Cell Death ,Autophagy ,Lysosome-Associated Membrane Glycoproteins ,Cell Biology ,Biology ,Cell biology ,Neoplasm Proteins ,Pathogenesis ,Cytoplasm ,Apoptosis ,Gene expression ,Humans ,Lysosomes ,Molecular Biology ,Research Paper - Abstract
Sjögren syndrome (SS) is a chronic and progressive autoimmune disease characterized by dry mouth and dry eyes, and characteristic autoantibodies. Evidence of altered macroautophagy/autophagy and apoptosis has been associated with SS, but a mechanistic understanding of the gene expression changes associated with these abnormal processes has not been realized. Recently, increased LAMP3 (lysosomal associated membrane protein 3) expression was found in a subset of SS patients and was associated with increased apoptosis and autoantigen accumulation and release. To better understand how LAMP3 expression might modulate apoptosis, cell biology, and biochemical studies were used to examine the effect of LAMP3 expression in minor salivary gland cells. LAMP3 expression resulted in degradation of LAMP1 increasing lysosomal membrane permeabilization and relocalization of cathepsins to the cytoplasm, resulting in destabilizing autophagic flux and caspase activation. These findings highlight the central role of LAMP3 expression in the pathogenesis of SS. ABBREVIATIONS: A253-control: A253 control for LAMP3 stable overexpression; A253- LAMP3: A253 LAPM3 stable overexpression; CASP1: caspase 1; CASP3: caspase 3; CHX: cycloheximide; CTSB: cathepsin B; CTSD: cathepsin D; CQ: chloroquine; DCs: dendritic cells; ER: endoplasmic reticulum; LGALS3: galectin 3; HCV: hepatitis C virus; HSG-control: HSG control for LAMP3 stable overexpression; HSG-LAMP3: HSG LAMP3 stable overexpression; HSP: heat shock protein; HTLV-1: human T-lymphocyte leukemia virus-1; IXA: ixazomib; LAMP: lysosomal associated membrane protein; MHC: major histocompatibility complex; mAb: monoclonal antibody; OE: overexpression; pepA: pepstatin A; pAb: polyclonal antibody; pSS: primary Sjögren syndrome; qRT-PCR: quantitative real- time reverse transcriptase polymerase chain reaction; SLE: systemic lupus erythematosus; SS: Sjögren syndrome; UPR: unfolded protein response; V-ATPase: vacuolar-type proton- translocating ATPase; Y-VAD: Ac-YVAD-cmk; Z-DEVD; Z-DEVD-fmk; Z-VAD: Z-VAD- fmk
- Published
- 2021
36. <scp>SARS CoV</scp>‐2 aggravates cellular metabolism mediated complications in<scp>COVID</scp>‐19 infection
- Author
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Yogendra Singh, Poonam Negi, Dinesh Kumar Chellappan, Imran Kazmi, Fahad A. Al-Abbasi, Gaurav Gupta, and Kamal Dua
- Subjects
Programmed cell death ,Coronavirus disease 2019 (COVID-19) ,Homocysteine ,viruses ,Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ,Dermatology ,Virus ,Short Papers ,030207 dermatology & venereal diseases ,03 medical and health sciences ,chemistry.chemical_compound ,Folic Acid ,0302 clinical medicine ,Ferroptosis ,Humans ,Short Paper ,Medicine ,Vitamin B12 ,Respiratory system ,Cellular metabolism ,Cell Death ,SARS-CoV-2 ,business.industry ,COVID-19 ,General Medicine ,Vitamin B 6 ,Vitamin B 12 ,chemistry ,030220 oncology & carcinogenesis ,Immunology ,business - Abstract
Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), the primary causative organism in corona virus disease-19 (COVID-19) infections, is a novel member of the human coronavirus family which was first identified in Wuhan, China, towards the end of 2019. This letter reveals new vital missing links in our current understanding of the mechanisms that lead to cell death triggered by ferroptotic stress in COVID-19 infection. It further reveal the importance of homocysteine mediated trans-sulfuration pathway in COVID-19 infection. Hence, Vitamin B6, folic acid, and Vitamin B12 should be incorporated in the treatment regimen for SARS CoV-2 infections to suppress complications, as the virus mediates altered host cell metabolism.
- Published
- 2020
37. RPW8.1 enhances the ethylene-signaling pathway to feedback-attenuate its mediated cell death and disease resistance in Arabidopsis
- Author
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Xiao-Rong He, Jing-Hao Zhao, Ji-Qun Zhao, Peng-Qiang Liu, Shunyuan Xiao, Zhi-Xue Zhao, Qin Feng, Wenming Wang, Ling-Li Zhang, Yan-Yan Huang, Yong-Ju Xu, Yan Li, and Jing Fan
- Subjects
0106 biological sciences ,0301 basic medicine ,ACO4 ,Programmed cell death ,Ethylene ,Physiology ,Arabidopsis ,Plant Science ,Plant disease resistance ,01 natural sciences ,Feedback ,03 medical and health sciences ,chemistry.chemical_compound ,Ascomycota ,Gene Expression Regulation, Plant ,ERF6 ,ORA59 ,EIN3/EIL1 ,Loss function ,Disease Resistance ,Plant Diseases ,RPW8.1 ,ethylene signaling ,Oxidase test ,biology ,Cell Death ,Full Paper ,Chemistry ,Arabidopsis Proteins ,Research ,Ethylene signaling pathway ,Ethylenes ,Full Papers ,biology.organism_classification ,Plants, Genetically Modified ,Cell biology ,030104 developmental biology ,Regulatory circuit ,powdery mildew ,ERF016 ,010606 plant biology & botany ,Signal Transduction - Abstract
Summary The Arabidopsis RESISTANCE TO POWDERY MILDEW 8.1 (RPW8.1) activates confined cell death and defense against different pathogens. However, the underlying regulatory mechanisms still remain elusive.Here, we show that RPW8.1 activates ethylene signaling that, in turn, negatively regulates RPW8.1 expression. RPW8.1 binds and stabilizes 1‐aminocyclopropane‐1‐carboxylate oxidase 4 (ACO4), which may in part explain increased ethylene production and signaling in RPW8.1‐expressing plants. In return, ACO4 and other key components of ethylene signaling negatively regulate RPW8.1‐mediated cell death and disease resistance via suppressing RPW8.1 expression.Loss of function in ACO4, EIN2, EIN3 EIL1, ERF6, ERF016 or ORA59 increases RPW8.1‐mediated cell death and defense response. By contrast, overexpression of EIN3 abolishes or significantly compromises RPW8.1‐mediated cell death and disease resistance. Furthermore, ERF6, ERF016 and ORA59 appear to act as trans‐repressors of RPW8.1, with OAR59 being able to directly bind to the RPW8.1 promoter.Taken together, our results have revealed a feedback regulatory circuit connecting RPW8.1 and the ethylene‐signaling pathway, in which RPW8.1 enhances ethylene signaling, and the latter, in return, negatively regulates RPW8.1‐mediated cell death and defense response via suppressing RPW8.1 expression to attenuate its defense activity.
- Published
- 2020
38. TNF-α-dependent neuronal necroptosis regulated in Alzheimer's disease by coordination of RIPK1-p62 complex with autophagic UVRAG
- Author
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Zhichu Ren, Ji Heon Noh, Evandro Fei Fang, Libin Wei, Jialin Wu, Yong U Liu, Jian Sima, Yuyuan Yao, Xijing Chen, Yiqun Wu, Chong Xu, and Guobing Chen
- Subjects
Necroptosis ,Medicine (miscellaneous) ,Gene Expression ,UVRAG ,Apoptosis ,Biology ,autophagic flux ,TNF-α/TNFR1 signaling ,RIPK1 ,Mice ,Necrosis ,RIPK1/RIPK3/MLKL cascade ,Downregulation and upregulation ,Alzheimer Disease ,Autophagy ,Animals ,Humans ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,Neurons ,Gene knockdown ,neuronal necroptosis ,Cell Death ,Tumor Necrosis Factor-alpha ,Tumor Suppressor Proteins ,p62 ,Brain ,RNA-Binding Proteins ,Alzheimer's disease ,Cell biology ,Disease Models, Animal ,Receptor-Interacting Protein Serine-Threonine Kinases ,Signal transduction ,Apoptosis Regulatory Proteins ,Transcriptome ,Necroptotic signaling pathway ,Signal Transduction ,Research Paper - Abstract
Background: Neuronal death is a major hallmark of Alzheimer's disease (AD). Necroptosis, as a programmed necrotic process, is activated in AD. However, what signals and factors initiate necroptosis in AD is largely unknown. Methods: We examined the expression levels of critical molecules in necroptotic signaling pathway by immunohistochemistry (IHC) staining and immunoblotting using brain tissues from AD patients and AD mouse models of APP/PS1 and 5×FAD. We performed brain stereotaxic injection with recombinant TNF-α, anti-TNFR1 neutralizing antibody or AAV-mediated gene expression and knockdown in APP/PS1 mice. For in vitro studies, we used TNF-α combined with zVAD-fmk and Smac mimetic to establish neuronal necroptosis models and utilized pharmacological or molecular biological approaches to study the signaling pathways. Results: We find that activated neuronal necroptosis is dependent on upstream TNF-α/TNFR1 signaling in both neuronal cell cultures and AD mouse models. Upon TNF-α stimulation, accumulated p62 recruits RIPK1 and induces its self-oligomerization, and activates downstream RIPK1/RIPK3/MLKL cascade, leading to neuronal necroptosis. Ectopic accumulation of p62 is caused by impaired autophagy flux, which is mediated by UVRAG downregulation during the TNF-α-promoted necroptosis. Notably, UVRAG overexpression inhibits neuronal necroptosis in cell and mouse models of AD. Conclusions: We identify a finely controlled regulation of neuronal necroptosis in AD by coordinated TNF-α signaling, RIPK1/3 activity and autophagy machinery. Strategies that could fine-tune necroptosis and autophagy may bring in promising therapeutics for AD.
- Published
- 2021
39. Corrigendum: FGF signaling regulates development by processes beyond canonical pathways
- Author
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Philippe Soriano, Pierre Mazot, Ayan Ray, J. Richard Brewer, Catarina Catela, and Colin J. Dinsmore
- Subjects
Cell Death ,Gene Expression Regulation, Developmental ,Computational biology ,Biology ,Fibroblast growth factor ,Corrigenda ,Receptors, Fibroblast Growth Factor ,Fibroblast Growth Factors ,Mice ,Neural Crest ,Mutation ,Cell Adhesion ,Genetics ,Animals ,Receptor, Fibroblast Growth Factor, Type 1 ,Receptor, Fibroblast Growth Factor, Type 2 ,Protein Kinases ,Cells, Cultured ,Signal Transduction ,Developmental Biology ,Research Paper - Abstract
FGFs are key developmental regulators that engage a signal transduction cascade through receptor tyrosine kinases, prominently engaging ERK1/2 but also other pathways. However, it remains unknown whether all FGF activities depend on this canonical signal transduction cascade. To address this question, we generated allelic series of knock-in Fgfr1 and Fgfr2 mouse strains, carrying point mutations that disrupt binding of signaling effectors, and a kinase dead allele of Fgfr2 that broadly phenocopies the null mutant. When interrogated in cranial neural crest cells, we identified discrete functions for signaling pathways in specific craniofacial contexts, but point mutations, even when combined, failed to recapitulate the single or double null mutant phenotypes. Furthermore, the signaling mutations abrogated established FGF-induced signal transduction pathways, yet FGF functions such as cell–matrix and cell–cell adhesion remained unaffected, though these activities did require FGFR kinase activity. Our studies establish combinatorial roles of Fgfr1 and Fgfr2 in development and uncouple novel FGFR kinase-dependent cell adhesion properties from canonical intracellular signaling.
- Published
- 2021
40. Computational modeling of therapy on pancreatic cancer in its early stages
- Author
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J. Chen, Fred J. Vermolen, and Daphne Weihs
- Subjects
Cancer therapy ,0206 medical engineering ,Cell ,Hyaluronoglucosaminidase ,02 engineering and technology ,Deoxycytidine ,Monte Carlo simulations ,Injections ,03 medical and health sciences ,Stroma ,Cell Movement ,Cell Line, Tumor ,Pancreatic cancer ,medicine ,Humans ,Cell migration ,Computer Simulation ,Neoplasm Staging ,030304 developmental biology ,Original Paper ,Stochastic Processes ,0303 health sciences ,Tumor microenvironment ,Cell Death ,business.industry ,Mechanical Engineering ,Cancer ,Numerical Analysis, Computer-Assisted ,medicine.disease ,Gemcitabine ,020601 biomedical engineering ,Extracellular Matrix ,Pancreatic Neoplasms ,medicine.anatomical_structure ,Modeling and Simulation ,Cancer cell ,Cancer research ,Anisotropy ,Mathematical modeling ,business ,Monte Carlo Method ,Cell Division ,Biotechnology ,medicine.drug - Abstract
More than eighty percent of pancreatic cancer involves ductal adenocarcinoma with an abundant desmoplastic extracellular matrix surrounding the solid tumor entity. This aberrant tumor microenvironment facilitates a strong resistance of pancreatic cancer to medication. Although various therapeutic strategies have been reported to be effective in mice with pancreatic cancer, they still need to be tested quantitatively in wider animal-based experiments before being applied as therapies. To aid the design of experiments, we develop a cell-based mathematical model to describe cancer progression under therapy with a specific application to pancreatic cancer. The displacement of cells is simulated by solving a large system of stochastic differential equations with the Euler–Maruyama method. We consider treatment with the PEGylated drug PEGPH20 that breaks down hyaluronan in desmoplastic stroma followed by administration of the chemotherapy drug gemcitabine to inhibit the proliferation of cancer cells. Modeling the effects of PEGPH20 + gemcitabine concentrations is based on Green’s fundamental solutions of the reaction–diffusion equation. Moreover, Monte Carlo simulations are performed to quantitatively investigate uncertainties in the input parameters as well as predictions for the likelihood of success of cancer therapy. Our simplified model is able to simulate cancer progression and evaluate treatments to inhibit the progression of cancer. Electronic supplementary material The online version of this article (10.1007/s10237-019-01219-0) contains supplementary material, which is available to authorized users.
- Published
- 2019
41. Mibefradil, a T‐type Ca 2+ channel blocker also blocks Orai channels by action at the extracellular surface
- Author
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Nawel Zaibi, Gui-Lan Chen, Thomas Hallett, Bo Zeng, Shang-Zhong Xu, Rahul Saurabh, Pengyun Li, and Hussein N. Rubaiy
- Subjects
0301 basic medicine ,Thapsigargin ,ORAI1 Protein ,ORAI2 Protein ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,medicine ,Extracellular ,Humans ,Channel blocker ,Patch clamp ,Cells, Cultured ,Cell Proliferation ,Pharmacology ,Mibefradil ,Cell Death ,ORAI1 ,Optical Imaging ,STIM1 ,Calcium Channel Blockers ,Research Papers ,3. Good health ,HEK293 Cells ,030104 developmental biology ,chemistry ,Biophysics ,Calcium ,Calcium Channels ,030217 neurology & neurosurgery ,Intracellular ,medicine.drug - Abstract
BACKGROUND AND PURPOSE: Mibefradil, a T‐type Ca(2+) channel blocker, has been investigated for treating solid tumours. However, its underlying mechanisms are still unclear. Here, we have investigated the pharmacological actions of mibefradil on Orai store‐operated Ca(2+) channels. EXPERIMENTAL APPROACH: Human Orai1‐3 cDNAs in tetracycline‐regulated pcDNA4/TO vectors were transfected into HEK293 T‐REx cells with stromal interaction molecule 1 (STIM1) stable expression. The Orai currents were recorded by whole‐cell and excised‐membrane patch clamp. Ca(2+) influx or release was measured by Fura‐PE3/AM. Cell growth and death were monitored by WST‐1, LDH assays and flow cytometry. KEY RESULTS: Mibefradil inhibited Orai1, Orai2, and Orai3 currents dose‐dependently. The IC(50) for Orai1, Orai2, and Orai3 channels was 52.6, 14.1, and 3.8 μM respectively. Outside‐out patch demonstrated that perfusion of 10‐μM mibefradil to the extracellular surface completely blocked Orai3 currents and single channel activity evoked by 2‐APB. Intracellular application of mibefradil did not alter Orai3 channel activity. Mibefradil at higher concentrations (>50 μM) inhibited Ca(2+) release but had no effect on cytosolic STIM1 translocation evoked by thapsigargin. Inhibition on Orai channels by mibefradil was structure‐related, as other T‐type Ca(2+) channel blockers with different structures, such as ethosuximide and ML218, had no or minimal effects on Orai channels. Moreover, mibefradil inhibited cell proliferation, induced apoptosis, and arrested cell cycle progression. CONCLUSIONS AND IMPLICATIONS: Mibefradil is a potent cell surface blocker of Orai channels, demonstrating a new pharmacological action of this compound in regulating cell growth and death, which could be relevant to its anti‐cancer activity.
- Published
- 2019
42. Conserved roles of glucose in suppressing reactive oxygen species-induced cell death and animal survival
- Author
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Yuehua Wei, Yinan Zhang, Congrong Wang, and Fengwen Li
- Subjects
Paraquat ,Aging ,Programmed cell death ,Longevity ,Mutant ,SKN-1/Nrf2 ,reactive oxygen species (ROS) ,Models, Biological ,chemistry.chemical_compound ,Species Specificity ,Diabetes mellitus ,medicine ,Animals ,Humans ,Caenorhabditis elegans ,Caenorhabditis elegans Proteins ,Transcription factor ,chemistry.chemical_classification ,Reactive oxygen species ,Cell Death ,Herbicides ,Chemistry ,Cell Biology ,Metabolism ,Carbohydrate ,medicine.disease ,Cell biology ,DNA-Binding Proteins ,Oxidative Stress ,Glucose ,hyperglycemia ,Reactive Oxygen Species ,Transcription Factors ,Research Paper - Abstract
Carbohydrate overconsumption increases blood glucose levels, which contributes to the development of various diseases including obesity and diabetes. It is generally believed that high glucose metabolism increases cellular reactive oxygen species (ROS) levels, damages insulin-secreting cells and leads to age-associated diabetic phenotypes. Here we find that in contrast, high glucose suppresses ROS production induced by paraquat in both mammalian cells and the round worm C. elegans. The role of glucose in suppressing ROS is further supported by glucose’s ability to alleviate paraquat’s toxicity on C. elegans development. Consistently, we find that the ROS-regulated transcription factor SKN-1 is inactivated by glucose. As a result, the ROS/SKN-1-dependent lifespan extension observed in paraquat-treated animals, mitochondrial respiration mutant isp-1 and germline-less mutant glp-1 are all suppressed by glucose. Our study reveals an unprecedented interaction of glucose with ROS, which could have significant impact on our current understanding of glucose- and ROS-related diseases.
- Published
- 2019
43. Apoptosis induced by Ibaraki virus does not affect virus replication and cell death in hamster lung HmLu-1 cells
- Author
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Rie Watanabe, Yuya Tsuruta, Hiroyuki Iwata, and Shusaku Shibutani
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Programmed cell death ,040301 veterinary sciences ,Ibaraki virus ,Hamster ,Arbovirus Infections ,Biology ,Virus Replication ,Arbovirus ,Cell Line ,0403 veterinary science ,03 medical and health sciences ,Virology ,Cricetinae ,medicine ,Animals ,Cytotoxicity ,Lung ,030304 developmental biology ,0303 health sciences ,Full Paper ,HmLu-1 ,IBAV ,Cell Death ,General Veterinary ,apoptosis ,04 agricultural and veterinary sciences ,medicine.disease ,Cell biology ,medicine.anatomical_structure ,Viral replication ,Apoptosis ,Arboviruses - Abstract
Ibaraki virus (IBAV) is an arbovirus that is transmitted by biting midges and causes Ibaraki disease in cattle. IBAV induces apoptosis in several mammalian cell lines, and apoptosis in turn facilitates IBAV replication. In addition, virus-induced apoptosis may contribute to mammalian-specific pathogenicity considering that some arboviruses induce apoptosis in mammalian cells but not in insect cells. In this study, we found that when hamster lung cells (HmLu-1) are used as a virus host, IBAV causes severe cytopathic effects with little induction of apoptosis. Furthermore, pharmacological inhibition of apoptosis did not affect IBAV-induced cytotoxicity. These results indicate the existence of an apoptosis-independent pathway in which IBAV replicates and exerts cytotoxicity in mammalian cells.
- Published
- 2019
44. Exosomes from mesenchymal stem cells modulate endoplasmic reticulum stress to protect against nucleus pulposus cell death and ameliorate intervertebral disc degeneration in vivo
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Yukun Zhang, Shengfeng Zhan, Yu Song, Cao Yang, Wenbin Hua, Kangcheng Zhao, Xinghuo Wu, Gaocai Li, Rongjin Luo, and Zhiwei Liao
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Adult ,Male ,0301 basic medicine ,Programmed cell death ,Nucleus Pulposus ,Adolescent ,Blotting, Western ,Fluorescent Antibody Technique ,Medicine (miscellaneous) ,Intervertebral Disc Degeneration ,exosomes ,Real-Time Polymerase Chain Reaction ,Young Adult ,03 medical and health sciences ,0302 clinical medicine ,In Situ Nick-End Labeling ,Animals ,Humans ,Medicine ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,Protein kinase B ,TUNEL assay ,Cell Death ,business.industry ,advanced glycation end products ,Endoplasmic reticulum ,Mesenchymal stem cell ,Mesenchymal Stem Cells ,Endoplasmic Reticulum Stress ,Microvesicles ,Rats ,030104 developmental biology ,Apoptosis ,030220 oncology & carcinogenesis ,Unfolded protein response ,Cancer research ,Female ,RNA Interference ,business ,Signal Transduction ,Research Paper - Abstract
Objectives: Intervertebral disc degeneration (IDD) is widely accepted as a cause of low back pain and related degenerative musculoskeletal disorders. Nucleus pulposus (NP) cell apoptosis which is related to excessive endoplasmic reticulum (ER) stress in the intervertebral disc (IVD) could aggravate IDD progression. Many studies have shown the therapeutic potential of exosomes derived from bone marrow mesenchymal stem cells (MSC-exos) in degenerative diseases. We hypothesized that the delivery of MSC-exos could modulate ER stress and inhibit excessive NP cell apoptosis during IDD. Methods: The ER stress levels were measured in normal or degenerative NP tissues for contrast. The effects of MSC-exos were testified in advanced glycation end products (AGEs) -induced ER stress in human NP cells. The mechanism involving AKT and ERK signaling pathways was investigated using RNA interference or signaling inhibitors. Histological or immunohistochemical analysis and TUNEL staining were used for evaluating MSC-exos therapeutic effects in vivo. Results: The ER stress level and apoptotic rate was elevated in degenerative IVD tissues. MSC-exos could attenuate ER stress-induced apoptosis by activating AKT and ERK signaling. Moreover, delivery of MSC-exos in vivo modulated ER stress-related apoptosis and retarded IDD progression in a rat tail model. Conclusions: These results highlight the therapeutic effects of exosomes in preventing IDD progression. Our work is the first to demonstrate that MSC-exos could modulate ER stress-induced apoptosis during AGEs-associated IVD degeneration.
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- 2019
45. Stimulation of TRPA1 attenuates ischemia-induced cardiomyocyte cell death through an eNOS-mediated mechanism
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Pritam Sinharoy, Spencer R. Andrei, Derek S. Damron, and Monica Ghosh
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Male ,0301 basic medicine ,Nitric Oxide Synthase Type III ,Biophysics ,cardiomyocytes ,Stimulation ,Nitric Oxide ,Endothelial NOS ,TRPA1 ,Biochemistry ,Nitric oxide ,Contractility ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Ischemia ,Enos ,Ca2+/calmodulin-dependent protein kinase ,Animals ,Humans ,Myocytes, Cardiac ,Phosphorylation ,TRPA1 Cation Channel ,Protein kinase B ,Cells, Cultured ,Mice, Knockout ,Cell Death ,biology ,Chemistry ,food and beverages ,biology.organism_classification ,Cell biology ,030104 developmental biology ,Calcium ,Proto-Oncogene Proteins c-akt ,psychological phenomena and processes ,030217 neurology & neurosurgery ,Research Paper ,Signal Transduction - Abstract
The functional expression of transient receptor potential cation channel of the ankyrin-1 subtype (TRPA1) has recently been identified in adult mouse cardiac tissue where stimulation of this ion channel leads to increases in adult mouse ventricular cardiomyocyte (CM) contractile function via a Ca2+-Calmodulin-dependent kinase (CaMKII) pathway. However, the extent to which TRPA1 induces nitric oxide (NO) production in CMs, and whether this signaling cascade mediates physiological or pathophysiological events in cardiac tissue remains elusive. Freshly isolated CMs from wild-type (WT) or TRPA1 knockout (TRPA1-/-) mouse hearts were treated with AITC (100 µM) and prepared for immunoblot, NO detection or ischemia protocols. Our findings demonstrate that TRPA1 stimulation with AITC results in phosphorylation of protein kinase B (Akt) and endothelial NOS (eNOS) concomitantly with NO production in a concentration- and time-dependent manner. Additionally, we found that TRPA1 induced increases in CM [Ca2+]i and contractility occur independently of Akt and eNOS activation mechanisms. Further analysis revealed that the presence and activation of TRPA1 promotes CM survival and viability following ischemic insult via a mechanism partially dependent upon eNOS. Therefore, activation of the TRPA1/Akt/eNOS pathway attenuates ischemia-induced CM cell death.
- Published
- 2019
46. Bacterial magnetosomes loaded with doxorubicin and transferrin improve targeted therapy of hepatocellular carcinoma
- Author
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Weiquan Liu, Jiaojiao Wang, Wei Jiang, Yunpeng Zhang, Abdul Basit, Xi Wang, Yuanyuan Geng, Ting Miao, and Junquan Liu
- Subjects
Male ,Carcinoma, Hepatocellular ,antitumor effect ,Iron ,medicine.medical_treatment ,Biomedical Engineering ,Mice, Nude ,Medicine (miscellaneous) ,Antineoplastic Agents ,Transferrin receptor ,doxorubicin ,Targeted therapy ,Circulating tumor cell ,In vivo ,Cell Line, Tumor ,Receptors, Transferrin ,polycyclic compounds ,medicine ,Animals ,Humans ,Doxorubicin ,Molecular Targeted Therapy ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,chemistry.chemical_classification ,Mice, Inbred BALB C ,Cell Death ,Chemistry ,Cell Membrane ,Liver Neoplasms ,digestive, oral, and skin physiology ,Transferrin ,technology, industry, and agriculture ,Bacterial magnetosomes ,hepatocellular carcinoma ,medicine.disease ,Endocytosis ,In vitro ,carbohydrates (lipids) ,stomatognathic diseases ,Hepatocellular carcinoma ,Cancer research ,Magnetosomes ,Research Paper ,Biotechnology ,medicine.drug - Abstract
High metastatic rate and recurrence of tumor because of tumor circulating cells are seriously hinders for clinical tumor therapy. Herein, we develop a novel, active-targeting nanotherapeutic by simultaneously loading doxorubicin (DOX) and transferrin (Tf) onto bacterial magnetosomes (Tf-BMs-DOX) and investigate its antitumor efficacy in vitro and in vivo. Drug release profiles indicated that Tf-BMs/BMs loaded with DOX were capable of sustained drug release, suggesting that reduce drugs required frequency of administration and enhance their therapeutic effect. The results of cellular uptake revealed that Tf-BMs-DOX recognized hepatocellular carcinoma HepG2 cells more specifically compared to HL-7702 normal hepatocytes because of high expression of transferrin receptor (TfR) on the surface of HepG2 cells. Tf-BMs-DOX increased tumor cytotoxicity and apoptosis more significantly than free DOX or BMs-DOX by regulating the expression of tumor-related and apoptosis-related genes. Following intravenous injection in HepG2 cell-bearing mice, Tf-BMs-DOX displayed tumor suppression rate of 56.78%, significantly higher than that of the BMs-DOX (41.53%) and free DOX (31.26%) groups. These results suggest that Tf-BMs-DOX have the potential to actively target to tumor sites, as well as the ability to kill circulating tumor cells via intravenous injection. Our findings provide a promising candidate for the clinical treatment of metastatic cancer.
- Published
- 2019
47. Overcoming EZH2 Inhibitor Resistance by Taxane in PTEN-Mutated Cancer
- Author
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Qiang Wu, R. Jeffrey Karnes, Haojie Huang, Yang Bai, Jun Zhang, Yunqian Pan, Linlin Ma, Yuqian Yan, Yinhui Yang, Xiaokun Gang, and Qiubo Lv
- Subjects
Male ,0301 basic medicine ,PTEN ,Tetrazolium Salts ,Medicine (miscellaneous) ,FOXO1 ,Mice, SCID ,taxane ,Mice ,Prostate cancer ,0302 clinical medicine ,Antineoplastic Combined Chemotherapy Protocols ,Gene expression ,Medicine ,Promoter Regions, Genetic ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,biology ,Forkhead Box Protein O1 ,EZH2 ,3. Good health ,cell death ,Docetaxel ,030220 oncology & carcinogenesis ,Taxoids ,Research Paper ,Protein Binding ,medicine.drug ,Bridged-Ring Compounds ,Mice, Nude ,Antineoplastic Agents ,macromolecular substances ,03 medical and health sciences ,Cell Line, Tumor ,cancer ,Animals ,Humans ,Enhancer of Zeste Homolog 2 Protein ,Taxane ,business.industry ,PTEN Phosphohydrolase ,Prostatic Neoplasms ,medicine.disease ,Thiazoles ,030104 developmental biology ,Drug Resistance, Neoplasm ,Mutation ,Cancer cell ,Cancer research ,biology.protein ,business - Abstract
Rationale: The Polycomb group (PcG) protein EZH2 is implicated in cancer progression due to its frequent overexpression in many cancer types and therefore is a promising therapeutic target. Forkhead box transcription factor-1 (FOXO1) is a tumor suppressor that is often transcriptionally downregulated in human cancers such as prostate cancer although the underlying regulatory mechanisms remain elusive. Methods: Analysis of EZH2 ChIP-seq and ChIP-on-chip data in various cell types was performed. ChIP-qPCR, RT-qPCR, and western blot analyses were conducted to determine the mechanism by which EZH2 represses FOXO1 expression. Immunohistochemistry was employed to assess the correlation between EZH2 and FOXO1 protein expression in prostate cancer patient specimens. In vitro MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) and animal experiments were performed to determine the anti-cancer efficacy of EZH2 inhibitor alone or in combination of docetaxel, a chemotherapy agent of the taxane family, and dependency of the efficacy on FOXO1 expression. Results: We demonstrated that EZH2 binds to the FOXO1 gene promoter. EZH2 represses FOXO1 gene expression at the transcriptional level. EZH2 protein level inversely correlated with FOXO1 protein expression in prostate cancer patient specimens. This repression requires the methyltransferase activity and the functional PRC2 complex. While effectively inducing loss of viability of PTEN-positive 22Rv1 prostate cancer cells, EZH2 inhibitor failed to inhibit growth of PTEN-negative C4-2 prostate cancer cells. Co-treatment with docetaxel overcame EZH2 inhibitor resistance in PTEN-negative cancer cells in vitro and in mice. This effect was largely mediated by docetaxel-induced nuclear localization and activation of FOXO1. Conclusions: This study identifies FOXO1 as a bona fide repression target of EZH2 and an essential mediator of EZH2 inhibition-induced cell death. Our findings suggest that EZH2 repression of FOXO1 can be targeted by EZH2 inhibitor as a monotherapy for PTEN-proficient cancers or in combination with taxane for treatment of cancers with PTEN mutation or deletion.
- Published
- 2019
48. Targeting and microenvironment-improving of phenylboronic acid-decorated soy protein nanoparticles with different sizes to tumor
- Author
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Weibo Cai, Cheng Li, Kangjun Yuan, Xiaoping Qian, Lei Ge, Rongshi Cheng, Xiqun Jiang, and Xu Zhen
- Subjects
Male ,Static Electricity ,phenylboronic acid ,Medicine (miscellaneous) ,02 engineering and technology ,010402 general chemistry ,01 natural sciences ,Mice ,chemistry.chemical_compound ,size effect ,In vivo ,Cell Line, Tumor ,Neoplasms ,Tumor Microenvironment ,medicine ,Animals ,Humans ,Tissue Distribution ,Doxorubicin ,Particle Size ,Phenylboronic acid ,Cytotoxicity ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,Soy protein ,Tumor microenvironment ,Cell Death ,Chemistry ,021001 nanoscience & nanotechnology ,Boronic Acids ,Endocytosis ,In vitro ,0104 chemical sciences ,Perfusion ,Drug Liberation ,Drug delivery ,Soybean Proteins ,Biophysics ,Nanoparticles ,0210 nano-technology ,drug delivery ,Research Paper ,medicine.drug - Abstract
It is essential for nanoparticles to delivery drugs accurately and penetrate deeply to tumor. However, complicated tumor microenvironment such as elevated tumor interstitial fluid pressure (IFP) and solid stress reduces the transport efficiency of nanomedicines in tumor. Methods: We herein report a drug delivery system of phenylboronic acid-decorated soy protein nanoparticles with the size of 30 nm, 50 nm and 150 nm. In vitro examinations including cytotoxicity, cellular uptake and penetration in multicellular tumor spheroids and in vivo observations including IFP and tumor solid stress measurements and antitumor activity were performed. Results: It was found that phenylboronic acid moiety could endow the nanoparticles actively targeting affinity to sialic acid (SA) which overexpressed in tumor cells. Simultaneously soy protein could improve tumor microenvironment such as reduction of IFP and tumor stress. Among the soy protein nanoparticles with different sizes, 30 nm-sized nanoparticles showed the best cellular uptake and highest cytotoxicity in vitro after loading doxorubicin (DOX). In vivo, 30 nm-sized nanoparticles showed the best tumor microenvironment improvement efficiency, leading to the enhanced drug accumulation and antitumor efficiency when combination with DOX. Conclusion: Our study introduces a bioactive nanoparticulate design strategy to actively target and significantly improve tumor microenvironment for enhanced cancer therapy.
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- 2019
49. The carbonic anhydrase IX inhibitor SLC-0111 sensitises cancer cells to conventional chemotherapy
- Author
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Francesca Bianchini, Fabrizio Carta, Lido Calorini, Claudiu T. Supuran, Silvia Peppicelli, Elena Andreucci, Jessica Ruzzolini, and Anna Laurenzana
- Subjects
Colorectal cancer ,medicine.medical_treatment ,chemotherapy ,01 natural sciences ,slc-0111 ,Antineoplastic Combined Chemotherapy Protocols ,Drug Discovery ,Tumor Cells, Cultured ,Carbonic Anhydrase Inhibitors ,Sulfonamides ,Cell Death ,Molecular Structure ,Melanoma ,General Medicine ,Dacarbazine ,Gene Expression Regulation, Neoplastic ,CAIX inhibitor ,Chemotherapy ,combined therapy ,drug resistance ,SLC-0111 ,Pharmacology ,Drug Discovery3003 Pharmaceutical Science ,MCF-7 Cells ,Fluorouracil ,Research Paper ,medicine.drug ,Structure-Activity Relationship ,Antigens, Neoplasm ,Temozolomide ,medicine ,Humans ,caix inhibitor ,Doxorubicin ,RNA, Messenger ,Carbonic Anhydrase IX ,Cell Proliferation ,Dose-Response Relationship, Drug ,010405 organic chemistry ,business.industry ,Phenylurea Compounds ,lcsh:RM1-950 ,Cancer ,HCT116 Cells ,medicine.disease ,0104 chemical sciences ,Radiation therapy ,010404 medicinal & biomolecular chemistry ,lcsh:Therapeutics. Pharmacology ,Cancer cell ,Cancer research ,Drug Screening Assays, Antitumor ,business - Abstract
Drug combination represents one of the most accredited strategies of cancer therapy able to improve drug efficacy and possibly overcome drug resistance. Among the agents used to complement conventional chemotherapy, carbonic anhydrase IX (CAIX) inhibitors appear as one of the most suitable, as markers of hypoxic and acidic cancer cells which do not respond to chemo- and radiotherapy. We performed preclinical in vitro assays to evaluate whether the SLC-0111 CAIX inhibitor co-operates and potentiates the cytotoxic effects of conventional chemotherapeutic drugs in A375-M6 melanoma cells, MCF7 breast cancer cells, and HCT116 colorectal cancer cells. Here, we demonstrate that the SLC-0111 CAIX inhibitor potentiates cytotoxicity of Dacarbazine and Temozolomide currently used for advanced melanoma treatment. SLC-0111 also increases breast cancer cell response to Doxorubicin and enhances 5-Fluorouracil cytostatic activity on colon cancer cells. These findings disclose the possibility to extend the use of CAIX inhibitors in the combination therapy of various cancer histotypes.
- Published
- 2018
50. AKT inhibition-mediated dephosphorylation of TFE3 promotes overactive autophagy independent of MTORC1 in cadmium-exposed bone mesenchymal stem cells
- Author
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Zhengping Yu, Huijuan Li, Min Yang, Chunhai Chen, Ping Deng, Mindi He, Huifeng Pi, Feng Gao, Zheng Wang, Menyu Liu, Zhou Zhou, Lingyun Zou, Tengfei Fan, Min Li, Jia Xie, Lei Zhang, Yonghui Lu, Mengyan Chen, Zhang Tao, Manyu Tu, Yu Xi, and Li Tian
- Subjects
0301 basic medicine ,Research Paper - Basic Science ,mTORC1 ,Mechanistic Target of Rapamycin Complex 1 ,Autophagy-Related Protein 7 ,Bone and Bones ,Mice ,03 medical and health sciences ,Sequestosome 1 ,Autophagy ,Animals ,Phosphorylation ,education ,Molecular Biology ,Protein kinase B ,Mechanistic target of rapamycin ,Cell Nucleus ,Microphthalmia-Associated Transcription Factor ,education.field_of_study ,Cell Death ,030102 biochemistry & molecular biology ,biology ,Basic Helix-Loop-Helix Leucine Zipper Transcription Factors ,Calcineurin ,Transcription factor EC ,Mesenchymal Stem Cells ,Cell Biology ,Cell biology ,Mice, Inbred C57BL ,030104 developmental biology ,Gene Expression Regulation ,FOXO3 ,biology.protein ,TFEB ,Lysosomes ,Proto-Oncogene Proteins c-akt ,Cadmium ,Signal Transduction - Abstract
Cadmium (Cd) is a toxic metal that is widely found in numerous environmental matrices and induces serious adverse effects in various organs and tissues. Bone tissue seems to be a crucial target of Cd contamination. Macroautophagy/autophagy has been proposed to play a pivotal role in Cd-mediated bone toxicity. However, the mechanisms that underlie Cd-induced autophagy are not yet completely understood. We demonstrated that Cd treatment increased autophagic flux and inhibition of the autophagic process using Atg7 gene silencing blocked the Cd-induced mesenchymal stem cell death. Mechanistically, Cd activated nuclear translocation of TFE3 but not that of TFEB or MITF, which contributed to the expression of autophagy-related genes and lysosomal biogenesis. Specifically, Cd decreased expression of phospho-AKT (Ser473). The reduction in AKT activity led to dephosphorylation of cytosolic TFE3 at Ser565 and promoted TFE3 nuclear translocation independently of MTORC1. Notably, Cd treatment increased the activity of PPP3/calcineurin, and pharmacological inhibition of PPP3/calcineurin with FK506 suppressed AKT dephosphorylation and TFE3 activity. These results suggest that PPP3/calcineurin negatively regulates AKT phosphorylation and is involved in Cd-induced TFE3-dependent autophagy. Modulation of the PPP3/calcineurin-AKT-TFE3 autophagic-lysosomal machinery may offer novel therapeutic approaches for the treatment of Cd-induced bone damage. Abbreviations: ACTB: actin: beta; AKT: thymoma viral proto-oncogene; AMPK: AMP-activated protein kinase; ATG: autophagy related; Baf A1: bafilomycin A(1); Cd: cadmium; FOXO3: forkhead box O3; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; MITF: melanogenesis associated transcription factor; MSC: mesenchymal stem sell; MTORC1: mechanistic target of rapamycin kinase complex 1; RPS6KB1: ribosomal protein S6 kinase: polypeptide 1; SGK1: serum/glucocorticoid regulated kinase 1; SQSTM1/p62: sequestosome 1;TFE3: transcription factor E3; TFEB: transcription factor EB; TFEC: transcription factor EC
- Published
- 2018
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