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Context: Current chemotherapeutic drugs cannot meet the treatment needs of patients with nasopharyngeal carcinoma (NPC), so urgent action is needed to discover novel chemotherapeutic agents. Our previous study revealed that garcinone E (GE) inhibited the proliferation and metastasis of NPC, suggesting that the compound might display promising anticancer activity., Objective: To examine the mechanism underlying the anti-NPC activity of GE for the first time., Materials and Methods: For MTS assay, NPC cells were treated with 2.5-20 μmol/L GE or dimethyl sulfoxide for 24, 48, and 72 h. Colony formation capacity, cell cycle distribution, and in vivo xenograft experiment of GE were assessed. MDC staining, StubRFP-sensGFP-LC3 observation, LysoBrite Blue staining, and immunofluorescence examined the autophagy of NPC cells after GE exposure. Western blotting, RNA-sequencing, and RT-qPCR measured protein and mRNA levels., Results: GE suppressed cell viability with an IC 50 of 7.64, 8.83 and 4.65 μmol/L for HK1, HONE1 and S18 cells. GE inhibited colony formation and cell cycle, increased autophagosome number, and inhibited the autophagic flux partially by blocking lysosome-autophagosome fusion, and repressed S18 xenograft growth. GE dysregulated the expression of autophagy- and cell cycle-related proteins such as Beclin-1, SQSTM1/p62, LC3, CDKs, and Cyclins. Bioinformatics GO and KEGG pathway enrichment analysis of RNA-seq showed that autophagy was enriched in differentially expressed genes upon GE treatment., Discussion and Conclusion: GE acts as an autophagic flux inhibitor, which may have potential chemotherapeutic use for NPC treatment and may have an application in basic research to explore the mechanisms of autophagy.

Tetrahydrocurcumin (THC) is an essential metabolite of curcumin, a major active component of the Curcuma species, which have been used traditionally for the treatment of cardiovascular diseases. The PI3K/AKT/mTOR signaling pathways serve a vital role during myocardial ischemia-reperfusion (MI/R) injury. The aim of the present study was to investigate the cardioprotective potential and mechanism of THC. In the in vivo study, an animal model of MI/R was induced by coronary occlusion. Results indicated that THC (50 mg/kg/day) protected the rat hearts from MI/R-induced heart failure by increasing ejection fraction (EF) and fractional shortening (FS) and decreasing left ventricular end systolic diameter (LVESD) and left ventricular end systolic volume (LVESV). THC also reduced myocardial infarct size and apoptosis. Furthermore, H9c2 cells were incubated with THC (20 μM) to explore its potential effect following exposure to hypoxia and reoxygenation (H/R). THC post-treatment significantly augmented cell viability and prevented lactate dehydrogenase (LDH) release after H/R exposure. THC effectively improved antioxidant activity by increasing SOD and CAT activities and decreasing MDA level. THC also enhanced mitochondrial membrane potential, inhibited apoptotic cell death, diminished the Bax/Bcl-2 ratio and cleaved caspase-3 level relative to the H/R model. In addition, THC effectively decreased Beclin1 expression and LC3 II/LC3 I ratio, but increased p62 expression, compared with the H/R model group, and decreased the formation of H/R-induced autophagosomes and autolysosomes. Furthermore, THC promoted the phosphorylation of PI3K/AKT/mTOR and induced the expression of hypoxia-inducible factor 1α (HIF-1α) after H/R. However, these effects on H9c2 cells were notably abolished by the PI3K inhibitor LY294002 and mTOR inhibitor rapamycin. In conclusion, THC effectively inhibited H/R-induced autophagy and apoptosis via, at least partially, activating the PI3K/AKT/mTOR pathways. THC might have the potential to be further developed into a potential candidate for the treatment of MI/R injury., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background/aims: Several miRNAs have been reported to be involved in the pathogenesis of polycystic ovarian syndrome (PCOS). However, the biological roles of miR-16 and its molecular mechanisms in PCOS development remain to be elucidated., Methods: qRT-PCR was performed to detect the expression levels of miR-16 and programmed cell death protein 4 (PDCD4). GCs proliferation, cell cycle distribution and apoptosis were examined by MTT assay and flow cytometry analysis. Luciferase reporter assay and RIP assay were applied to confirm the regulatory relationship between miR-16 and PDCD4. Western blot was applied to measure the protein levels of PDCD4, PCNA and caspase-3. ELISA kits were used to determine the serum levels of steroids., Results: miR-16 expression was down-regulated in ovarian cortex tissues and serums of PCOS patients. PDCD4 expression was up-regulated in ovarian cortex tissues of PCOS patients. miR-16 overexpression facilitated cell proliferation, induced cell cycle progression, and inhibited apoptosis in GCs. Moreover, PDCD4 was a direct target of miR-16. Also, enforced expression of PDCD4 abated the effects of miR-16 on GCs growth and apoptosis. Additionally, testosterone resulted in a decrease of miR-16 expression and an increase of PDCD4 expression, thus blocking cell growth and enhanced apoptosis in GCs. Furthermore, miR-16 overexpression alleviated PCOS in vivo by regulating PDCD4., Conclusions: miR-16 promoted ovarian GCs proliferation and inhibited apoptosis through directly targeting PDCD4 in PCOS, contributing to a better understanding of the molecular mechanism of GCs dysregulation and providing a promising target in PCOS., (© 2018 The Author(s). Published by S. Karger AG, Basel.)

Background: Chemotherapy-induced premature ovarian failure (POF) is a severe complication affecting tumor patients at a childbearing age. Mesenchymal stem cells (MSCs) can partially restore the ovarian structure and function damaged by chemotherapy. miR-21 is a microRNA that can regulate cell apoptosis. This study discusses the repair effect and mechanism of MSCs overexpressing miR-21 on chemotherapy-induced POF., Methods: Rat MSCs and granulosa cells (GCs) were isolated in vitro. MSCs were transfected with miR-21 lentiviral vector (LV-miR-21) to obtain MSCs stably expressing miR-21 (miR-21-MSCs). The microenvironment of an ovary receiving chemotherapy was mimicked by adding phosphamide mustard (PM) into the cellular culture medium. The apoptosis rate and the mRNA and protein expression of target genes PTEN and PDCD4 were detected in MSCs. Apoptosis was induced by adding PM into the culture medium for GCs, which were cocultured with miR-21-MSCs. The apoptosis rate and the mRNA and protein expression of PTEN and PDCD4 were detected. The chemotherapy-induced POF model was built into rats by intraperitoneal cyclophosphamide injection. miR-21-MSCs were transplanted into the bilateral ovary. The rats were sacrificed at 15, 30, 45, and 60 days after the last injection. The ovarian weights, follicle count, estrous cycle, and sex hormone levels (estradiol (E2) and follicle-stimulating hormone (FSH)) were detected. Apoptosis of GCs was determined by TUNEL assay. The miR-21 and mRNA and protein expression of PTEN and PDCD4 were determined., Results: The apoptosis decreased in MSCs transfected with miR-21. The mRNA and protein expression of target genes PTEN and PDCD4 was downregulated. GCs cocultured with miR-21-MSCs showed a decreased apoptosis, an upregulation of miR-21, and a downregulation of PTEN and PDCD4. Following the injection of miR-21-MSCs, the ovarian weight and follicle counts increased; E 2 levels increased while FSH levels decreased, with less severe apoptosis of GCs. The miR-21 expression in the ovaries was upregulated, while the mRNA expression and protein expression of PTEN and PDCD4 were downregulated., Conclusions: Overexpression of miR-21 in MSCs promoted efficacy against chemotherapy-induced POF and its improvement of the repair effect was related to the inhibition of GC apoptosis by targeting PTEN and PDCD4.

Sporamin, a Kunitz-type trypsin inhibitor (TI) from sweet potato tuberous roots, has demonstrated anti-tumor activity through poorly defined mechanisms. Furthermore, the effects of sporamin on pancreatic cancer have not been explored. Herein, we studied the effects of sporamin on two human pancreatic cancer cell lines, PANC-1 and BxPC-3. Sporamin significantly inhibited the cell viability and proliferation activity and induced apoptosis in PANC-1 and BxPC-3 cells. Consistently, in sporamin-treated PANC-1 and BxPC-3 cells, the anti-apoptotic proteins Bcl-2 and Bcl-XL were downregulated and the pro-apoptotic protein Bax was upregulated. Moreover, nuclear factor kappa B activation and IκBα phosphorylation were inhibited, and total IκBα expression was increased in sporamin-treated PANC-1 and BxPC-3 cells. Thus, our results suggest that the anti-tumor effects of sporamin in pancreatic cancer cells might result partly from induction of apoptosis by downregulating nuclear factor kappa B pathway.

Pulsed electromagnetic field (PEMF), a non-invasive physical treatment modality, is now used clinically to promote bone formation for osteoporosis. The patients after ectomy of ovarian cancer are easily complicated with osteoporosis. However, the safety parameters of PEMF treatment for the osteoporosis patients after resection of ovarian cancer remain unknown. Therefore, this study was designed to examine the effect of different frequency of PEMF on the proliferation, apoptosis and migration of human ovarian cancer cells (SKOV3 cells). Cultured SKOV3 cells were exposed to PEMF stimulation daily with radiation of 8 Hz, 16 Hz, 32 Hz and 64 Hz, respectively. We used sinusoidal waves with strength of 1 mT, twice a day with an interval of 12 hours. An exposure to the waves lasted 30 minutes, for 3 days, with those no PEMF stimulation serving as the control. The proliferation of cells was detected using EdU assay, and the apoptosis of cell was assessed with Annexin V-FITC fluorescence. The migration of cells was measured with the scratch wound assay. The data showed that exposure to PEMF of 1 mT, 8 Hz for 3 days could significantly inhibit the proliferation of SKOV3 cells and induce the apoptosis of the cells. The migrated distance and number were increased by 1 mT, 8 Hz or 32 Hz PEMF stimulation, but decreased by 1 mT, 16 Hz treatment. The results suggested that we should be careful about the safety of PEMF treatment and strictly choose the optical parameters in preventing or treating the osteoporosis of the patients after resection of ovarian cancer.

Contaminants in food have severely threatened human health, and appropriate antioxidants derived from food could reduce impairment risk. Lactoferrin from milk could control iron concentration in the blood to ameliorate oxidative stress, which is also required for sperm maturation, but the underlying mechanisms remain unclear. The present study used mice with spermatogenetic dysfunction caused by bisphenol A (BPA) and cadmium (Cd) to evaluate the ameliorative effects of lactoferrin and milk (bioactive substances). BPA (50 mg/kg) and Cd (1.6 mg/kg) caused severe damage to testis, including globally decreased germ cell counts, poor sperm quality, disordered apoptosis, oxidative stress, and autophagy; however bioactive substances comprehensively ameliorated spermatogenetic dysfunction via mitigating the increased levels of BAX/BCL2, LC3II/LC3I, and P62. AMPK was involved in autophagic regulation, while ERK1/2 inhibition attenuated the protective effects of lactoferrin, including restimulating apoptosis, oxidative stress, and arrested autophagic flux. Notably, P62 was consistently stimulated with different ERK1/2 inhibitors, which was ubiquitin-dependent. The study provides evidence for the alleviative effects of lactoferrin and milk in mice with spermatogenetic dysfunction through ERK1/2 mediated the ubiquitin-dependent degradation of P62. The involved signals and molecules could be identified as novel therapeutic targets for male infertility, which contributes to expanding LF's interests in research and application. [Display omitted] • Milk and LF protect mice from testicular toxicity induced by contaminants. • Milk and LF maintain disordered apoptosis, oxidative stress, and autophagy. • AMPK mainly participats in the remission process of autophagic disorder. • ERK1/2 plays critical roles in apoptosis, oxidative stress, and autophagy. • P62 plays crucial roles in testicular toxicity via ubiquitin-mediated degradation. [ABSTRACT FROM AUTHOR]

Background: Breast cancer (BC) is a prevalent female cancer, which has high morbidity and mortality. However, the pathogenesis of BC has not been fully elucidated. Studies have shown that TGF-β1 plays an important role in regulating the balance between autophagy and apoptosis of tumor. We aim to clarify the specific mechanism of autophagy and apoptosis in breast cancer maintaining the tumor microenvironment. Methods: The clinical characteristics of 850 BC patients were retrieved from the TCGA database. Differentially expressed autophagy-related genes (DEARGs) between tumor and normal tissues were obtained by the Wilcox test. Through Cox proportional hazard regression analysis, the prognostic risk model was constructed and verified by the ROC curve. We used MDC staining, colony formation assay, CCK-8, flow cytometric analysis to confirm the importance of TGF-β1 on the autophagy and apoptosis of breast cancer cells. Furthermore, western blot was performed to determine the relative expression of protein. The Kaplan-Meier Plotter database was utilized to identify the prognostic value of TP63. Results: We successfully constructed a prognostic risk model of breast cancer and screened out an autophagy-related prognostic gene -TP63. We predicted that TGF-β1 and TP63 have a binding site in the JASPAR database as expected. Additionally, TGF-β1 promoted autophagy and inhibited apoptosis of breast cancer cells by inhibiting the expression of TP63. Conclusion: Our study demonstrated that the molecular mechanism of TGF-β/TP63 signaling in regulating autophagy and apoptosis of breast cancer and provided a potential prognostic marker in breast cancer. [ABSTRACT FROM AUTHOR]

The prescription of Yiqi Jiedu decoction (YQJD) originated from the classic Chinese herbal prescriptions of Danggui Buxue Decoction and Wuzi Yanzong Pill. A previous study has shown that 4 Gy irradiation induced the apoptosis of spermatocytes and revealed autophagosomes in cells exposed to radiation. YQJD decoction has the effect of preventing radiation injury. We used spermatocytes (GC-2spd cell line) to investigate the relationship between autophagy and apoptosis of spermatogenic cells after radiation, and the mechanisms of YQJD decoction. Establish an in vitro radiation injury model by irradiating GC-2spd cells with 60Co γ-rays (4 Gy or 8 Gy). Autophagy agonists, autophagy inhibitors and YQJD were used to intervene cells. Cell apoptosis and inflammatory factors were measured. NF-κB localization was observed by immunofluorescence. Autophagy and apoptosis-related proteins and IκBα/NF-κB pathway factors were detected. Ionizing radiation promoted the growth of spermatogenic autophagosomes. After radiation, NF-κB was translocated to the nucleus, inflammatory factors were secreted, and IκBα/NF-κB pathway was activated, which promoted autophagy and apoptosis. YQJD decoction can inhibit the phosphorylation of IκBα/NF-κB pathway related factors, regulate the expression of Beclin-1 and Bcl-2 proteins, and inhibit the occurrence of autophagy and apoptosis of irradiated spermatocyte. The research results indicate that ionizing radiation can activate the IκBα/NF-κB signaling pathway in spermatocytes, promote cell autophagy and apoptosis by regulating the expression of Beclin-1 and Bcl-2 factors. The YQJD decoction inhibits the IκBα/NF-κB signaling pathway so as to regulate Beclin-1 and Bcl-2. [Display omitted] • Increased irradiation dose aggravated the blocking of autophagy flux. • Ionizing radiation promotes the binding of p65 to the Beclin-1 binding site, and therefore induce autophagy and apoptosis. • YQJD decoction prevents spermatocytes radiation injury via suppressing IκBα/NF-κB pathway. [ABSTRACT FROM AUTHOR]

The aim of the present study was to investigate the effects of daphnetin combined with B cell lymphoma 2 (Bcl2)-targeted small interfering (si)RNA (si-Bcl2) on antiapoptotic genes in fibroblast-like synoviocytes (FLS) in rats with collagen II-induced arthritis (CIA). The roles of si-Bcl2 and daphnetin were determined by measuring the expression levels of Bcl2. Protein and mRNA expression levels of Bcl2 in FLS were determined by flow cytometry and reverse transcription-quantitative polymerase chain reaction. Apoptosis of FLS was also determined by flow cytometry. It was revealed that treatment with si-Bcl2 or daphnetin alone resulted in downregulation of Bcl2 mRNA and protein expression. In addition, the mRNA expression levels of the signal transducer and activator of transcription 3 (STAT3), which transcriptionally regulates the activity of mitochondria, were reduced. The combination of si-Bcl2 and daphnetin exhibited an enhanced effect on rheumatoid arthritis FLS (RAFLS), in which the apoptotic rate was significantly higher than either treatment alone. The results suggested that si-Bcl combined with daphnetin may have an enhanced effect in promoting apoptosis of RAFLS derived from CIA rats, and a possible underlying molecular mechanism may function through the downregulation of Bcl2 expression and STAT3, which is located upstream of Bcl2 in the mitochondrial apoptotic pathway. The results of the present study are expected to provide theoretical and experimental basis for the treatment of RA and the medicinal development of daphnetin combined siRNA. [ABSTRACT FROM AUTHOR]

Objective CKLF-like MARVEL transmembrane domain containing 3 ( CMTM3 ), as a tumor suppressor gene, plays an important role in the suppression of cell growth and apoptosis. The goal of our study is to investigate the association between CMTM3 promoter methylation and laryngeal squamous cell carcinoma (LSCC). Design and methods Using the bisulfite pyrosequencing technology, DNA methylation levels of seven CpG sites in CMTM3 promoter are measured in tumor tissues and their adjacent tissues of 76 male LSCC patients. Results Our results reveal a significantly elevated promoter methylation of CMTM3 in tumor tissues compared with their adjacent tissues ( P < 0.001). A breakdown analysis by age shows that significant association of CMTM3 promoter methylation with cancer risk is specific to the LSCC patients older than 55 years ( P < 0.001) but not in the younger patients ( P = 0.305). Moreover, the association is only observed in the LSCC patients with smoking behavior ( P = 0.001). Breakdown analysis also shows that CMTM3 promoter methylation is associated with cancer risk among patients with stage I LSCC ( P < 0.001). Conclusion In conclusion, our study indicates that elevated CMTM3 methylation is a risk factor in male LSCC patients, especially in the patients with age over 55 years and with smoking behavior. [ABSTRACT FROM AUTHOR]

Ionizing radiation (IR) has found widespread application in modern medicine. As a result, radiotherapy inevitably causes spermatogenic cell injury. Many Chinese herbal prescriptions or natural extracts have the potential to protect against radiation injury. We used GC-2spd cells to investigate the effects and potential mechanisms of YQJD decoction on protecting spermatogenic cells from ionizing radiation injury. Firstly, the GC-2spd cells were irradiated with 60Co γ-rays (1 Gy, 2 Gy, 4 Gy and 8 Gy) to establish an in vitro model of radiation injury. After that, Cells were divided into six groups: negative control group (NC group), model group (IR group), positive drug group (IRA group), high-dose YQJD decoction (IRH group), medium-dose YQJD decoction (IRM group), and low-dose YQJD decoction group (IRL group). DNA damage, oxidative damage and inflammatory factors were measured. Cell apoptosis and cell cycle were detected by Flow cytometry. Transmission electron microscopy was performed to observe the morphological changes. After irradiation with 60CO γ-ray, the results indicated that the damage of spermatocyte was significantly induced by radiation exposure over 4 Gy. Furthermore, ionizing radiation could make DNA damage and oxidative stress in in GC-2spd cells. In addition, 60CO γ-ray also caused the increase of IL-1β, IL-6 and TNF-α and the change of cell cycle. However, the application of YQJD decoction inhibited the damage and apoptosis of GC-2spd cells in the aspects of anti-oxidation, promoting DNA damage repair and regulating inflammatory reaction. Taken together, the protective effects of YQJD decoction on 60CO γ-ray induced spermatocyte injury were confirmed in this study. This exploration might provide a new strategy for the application of Chinese herbs in radioprotection. [Display omitted] • Oxidative damage and inflammation induced by ionizing radiation hinder the repair of spermatogenic cells. • YQJD have the potential to alleviate DNA damage in spermatogenic cells. • The anti-oxidative and anti-inflammatory effects of YQJD reduce the indirect radiation damage to spermatogenic cells. [ABSTRACT FROM AUTHOR]

Silver nanoparticles (AgNPs) are pervasive environmental pollutants capable of inducing toxicological impacts on benthic organisms. In this study, the effects of AgNPs on the antioxidant enzyme activities, tissue damage, inflammatory responses, and reproductive toxicity of Corbicula fluminea were investigated. C. fluminea was exposed to four concentrations of AgNPs (0, 5 mg/L, 10 mg/L, and 125 mg/L) for 48 h. The results showed that the higher concentrations of AgNPs caused severe tissue damage in multiple organs of C. fluminea , induced oxidative stress and an imbalance of the antioxidant enzyme activities (such as SOD, CAT, MDA), and increased the inflammatory immune response involving NFκB, TLR2/4, HSP70/90, IL1β, and TNFα. Notably, further transmission electron microscopy and cytological analyses revealed that AgNPs exposure induced apoptosis in the gonad tissues, resulting in significant loss and damage in the oocytes and spermatids. The present study demonstrates the ecotoxicological impacts of AgNPs on freshwater bivalves, particularly highlighting their reproductive toxicity on germ cells, signifying the potential toxic effects of heavy metal pollution on aquatic ecosystems. AgNPs induce Ag deposition in various tissues, histopathological alterations, oxidative stress, inflammatory response, and germ cell apoptosis, resulting in significant toxic effects in Asian Clam (Corbicula fluminea). [Display omitted] • AgNPs exposure caused deposition of silver particles in tissues of Corbicula fluminea and lead to structural damage. • AgNPs exposure induced oxidative stress and impaired the activity of antioxidant enzymes in Corbicula fluminea tissues. • AgNPs exposure triggered immune responses. • AgNPs exposure induced reproductive toxicity and leads to apoptosis in gonad tissues. [ABSTRACT FROM AUTHOR]

Irradiation-induced immunosuppression often occurs during radiotherapy in patients, which would increase the risk of opportunistic infections. Many Chinese herbal prescriptions or natural extracts have recently attracted increased radiation protection and therapy attention due to their low toxicity. The present study aimed to investigate the protective effects of Yiqi Jiedu (YQJD) decoction on spleen injury induced by 2 Gy 60Co γ ray in mice. A total of 180 Balb/c mice were randomly divided into five groups: blank control (Ctrl), model (IR), positive drug (IRA), low-dose YQJD decoction (IRL), and high-dose YQJD decoction (IRH). After a ten-day intervention, mice were exposed to a single dose of total body irradiation (2 Gy) and sacrificed on the 1st, 3rd, and 7th day after irradiation. The indicators include general observations and body weight, changes in peripheral hemogram, index and histopathology examination of the spleen, distribution of lymphocyte subsets, cytokine levels, and apoptosis in the spleen. In comparison to the Ctrl group, the body weight, spleen index, peripheral blood cell, and splenocyte quantities decreased significantly after exposure, accompanied by a notable increase of apoptosis in spleen cells. Moreover, ionizing radiation also broke the balance of CD4+/CD8+, Th1/Th2, and Th17/Treg, triggering immune imbalance and immunosuppression. The above injuries occurred on the 1st day after exposure, worsened on the 3rd, and were relieved on the 7th day. However, the pretreatment of YQJD decoction increased the spleen index, improved the spleen structure, and inhibited radiation-induced apoptosis after exposure. Additionally, YQJD decoction has shown its ability to promote immunological balance recovery following exposure by regulating CD4+/CD8+, Th1/Th2, and Th17/Treg ratios, which may minimize the risk of infection. In addition, the high-dose of YQJD decoction showed a better protective effect than the low-dose group. The protective effects of YQJD decoction on 2 Gy 60Coγray induced spleen injury were confirmed in this study. This mechanism may be related to inhibiting apoptosis and modulating immune balance. This exploration might provide new insights into the use of Chinese herbs on radioprotection of the immune system. [Display omitted] • YQJD decoction effectively attenuated 2 Gy 60Co γ ray induced spleen injury. • YQJD decoction increased the spleen index, improved the spleen structure and inhibited apoptosis after exposure. • YQJD decoction promoted the recovery of immune balance by regulating CD4+/CD8+, Th1/Th2 and Th17/Treg after exposure. • Overall, YQJD decoction attenuated radiation-induced spleen injury by inhibiting apoptosis and modulating immune balance. [ABSTRACT FROM AUTHOR]