103 results on '"Rosa, Angela"'
Search Results
2. Role of rhBMP-7, Fibronectin, And Type I Collagen in Dental Implant Osseointegration Process: An Initial Pilot Study on Minipig Animals
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Gianmario Schierano, Roberto Navone, Giulio Preti, Bruno Peirone, Giuliana Muzio, Mitzy Mauthe von Degerfeld, and Rosa Angela Canuto
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Technology ,medicine.medical_treatment ,Inflammation ,Bone morphogenetic protein ,Osseointegration ,Article ,Andrology ,03 medical and health sciences ,0302 clinical medicine ,fibronectin ,medicine ,bone morphogenetic proteins ,type 1 collagen ,General Materials Science ,Dental implant ,030304 developmental biology ,0303 health sciences ,Microscopy ,QC120-168.85 ,dental implant ,biology ,business.industry ,QH201-278.5 ,rhBMP-7 ,030206 dentistry ,Engineering (General). Civil engineering (General) ,cytokines ,TK1-9971 ,Fibronectin ,Descriptive and experimental mechanics ,biology.protein ,Osteocalcin ,dental implant, fibronectin, rhBMP-7, type 1 collagen, cytokines, bone morphogenetic proteins ,Implant ,Electrical engineering. Electronics. Nuclear engineering ,medicine.symptom ,TA1-2040 ,business ,Type I collagen - Abstract
Background: The biological factors involved in dental implant osseointegration need to be investigated to improve implant success. Methods: Twenty-four implants were inserted into the tibias of six minipigs. Bone samples were obtained at 7, 14, and 56 days. Biomolecular analyses evaluated mRNA of BMP-4, -7, Transforming Growth Factor-β2, Interleukin-1β, and Osteocalcin in sites treated with rhBMP-7, Type 1 Collagen, or Fibronectin (FN). Inflammation and osteogenesis were evaluated by histological analyses. Results: At 7 and 14 days, BMP-4 and BMP-7 increased in the sites prepared with rhBMP-7 and FN. BMP-7 remained greater at 56 days in rhBMP-7 and FN sites. BPM-4 at 7 and 14 days increased in Type 1 Collagen sites, BMP-7 increased from day 14. FN increased the TGF-β2 at all experimental times, whilst the rhBMP-7 only did so up to 7 days. IL-1β increased only in collagen-treated sites from 14 days. Osteocalcin was high in FN-treated sites. Neutrophilic granulocytes characterized the inflammatory infiltrate at 7 days, and mononuclear cells at 14 and 56 days. Conclusions: This initial pilot study, in a novel way, evidenced that Type 1 Collagen induced inflammation and did not stimulate bone production, conversely FN or rhBMP-7 showed neo-osteogenetic and anti-inflammatory properties when directly added into implant bone site.
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- 2021
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3. Pathogenesis and Management of COVID-19
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Mona F. Shabana, Ali S Alqahtani, Heyam Saad Ali, Sari T S Alhoufie, Mohamed E. Ahmed, Ahmed Qasim Ahmed, Rosa Angela Cardone, AbdelRahman M. Ramadan, Samrein B M Ahmed, Ali Alqahtani, Laurent Schwartz, Saad S. Alqahtani, Muntaser E. Ibrahim, Adil H. H. Bashir, Khalid O. Alfarouk, Stephan J. Reshkin, and Jesús Devesa
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0301 basic medicine ,2019-20 coronavirus outbreak ,Coronavirus disease 2019 (COVID-19) ,Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ,RM1-950 ,Review ,SARS-COV-2 ,virus ,Biology ,Toxicology ,Virus ,Pathogenesis ,03 medical and health sciences ,0302 clinical medicine ,RA1190-1270 ,Pandemic ,medicine ,Cytokine expression ,COVID-19 ,medicine.disease ,Pollution ,030104 developmental biology ,030220 oncology & carcinogenesis ,Toxicology. Poisons ,cytokine storm ,Therapeutics. Pharmacology ,pharmacology ,Cytokine storm ,Neuroscience - Abstract
COVID-19, occurring due to SARS-COV-2 infection, is the most recent pandemic disease that has led to three million deaths at the time of writing. A great deal of effort has been directed towards altering the virus trajectory and/or managing the interactions of the virus with its subsequent targets in the human body; these interactions can lead to a chain reaction-like state manifested by a cytokine storm and progress to multiple organ failure. During cytokine storms the ratio of pro-inflammatory to anti-inflammatory mediators is generally increased, which contributes to the instigation of hyper-inflammation and confers advantages to the virus. Because cytokine expression patterns fluctuate from one person to another and even within the same person from one time to another, we suggest a road map of COVID-19 management using an individual approach instead of focusing on the blockbuster process (one treatment for most people, if not all). Here, we highlight the biology of the virus, study the interaction between the virus and humans, and present potential pharmacological and non-pharmacological modulators that might contribute to the global war against SARS-COV-2. We suggest an algorithmic roadmap to manage COVID-19.
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- 2021
4. A Small-Scale shRNA Screen in Primary Mouse Macrophages Identifies a Role for the Rab GTPase Rab1b in Controlling Salmonella Typhi Growth
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Virtu Solano-Collado, Rosa Angela Colamarino, David A. Calderwood, Massimiliano Baldassarre, and Stefania Spanò
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0301 basic medicine ,Microbiology (medical) ,Rab1b ,Immunology ,Population ,shRNA screen ,lcsh:QR1-502 ,GTPase ,Biology ,Salmonella typhi ,Microbiology ,complex mixtures ,Salmonella Typhi ,lcsh:Microbiology ,Small hairpin RNA ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,education ,Pathogen ,030304 developmental biology ,0303 health sciences ,education.field_of_study ,Intracellular parasite ,Rab GTPases ,bacterial infections and mycoses ,3. Good health ,macrophages ,030104 developmental biology ,Infectious Diseases ,host-defense ,bacteria ,Rab ,030217 neurology & neurosurgery - Abstract
Salmonella Typhi, the causative agent of typhoid fever, a life-threatening systemic infection, is a human restricted bacterial pathogen. A fundamental aspect of S. Typhi pathogenesis is its ability to survive in human macrophages but not in macrophages from other animals (i.e. mice). Despite the importance of macrophages in establishing systemic S. Typhi infection, the mechanisms that macrophages use to control the growth of S. Typhi and the role of these mechanisms in the bacterium’s adaptation to the human host are mostly unknown. To facilitate unbiased identification of genes involved in controlling the growth of S. Typhi in macrophages, we report optimized experimental conditions required to perform loss-of function pooled shRNA screens in primary mouse bone-marrow derived macrophages. Following infection with a fluorescently-labeled S. Typhi, cells defective in genes important for controlling S. Typhi growth (and therefore unable to kill S. Typhi) are sorted based on the intensity of fluorescence (i.e. number of intracellular fluorescent bacteria). shRNAs enriched in the fluorescent population are identified by next-generation sequencing. A proof-of-concept screen targeting the mouse Rab GTPases confirmed Rab32 as important to restrict S. Typhi in mouse macrophages. Interestingly and rather unexpectedly, this screen also revealed that Rab1b controls S. Typhi growth in mouse macrophages. This constitutes the first report of a Rab GTPase other than Rab32 involved in S. Typhi host-restriction. The methodology described here should allow genome-wide screening to identify mechanisms controlling the growth of S. Typhi and other intracellular pathogens in primary immune cells.
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- 2021
5. Phosphorylation of NHERF1 S279 and S301 differentially regulates breast cancer cell phenotype and metastatic organotropism
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Séverine Marionneau-Lambot, Emeline Bon, Stefania Cannone, Sébastien Roger, Thibauld Oullier, Roseline Guibon, Maria Raffaella Greco, Rosa Rubino, Rosa Angela Cardone, Manuel Bernabe-Garcia, Gaëlle Fromont, Maria-Luisa Cayuela, Stephan J. Reshkin, Loredana Ciaccia, and Lorenzo Guerra
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0301 basic medicine ,Scaffold protein ,Sodium-Hydrogen Exchangers ,PDZ domain ,Breast Neoplasms ,Biology ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Cell Line, Tumor ,Animals ,Humans ,Neoplasm Invasiveness ,Vasculogenic mimicry ,Neoplasm Metastasis ,Phosphorylation ,Molecular Biology ,Zebrafish ,Sodium-Hydrogen Exchanger 1 ,Hydrogen-Ion Concentration ,Phosphoproteins ,Xenograft Model Antitumor Assays ,Phenotype ,Cell biology ,Gene Expression Regulation, Neoplastic ,Disease Models, Animal ,030104 developmental biology ,030220 oncology & carcinogenesis ,Cancer cell ,Molecular Medicine ,Female ,Mutant Proteins ,Signal transduction ,Reprogramming ,Signal Transduction - Abstract
Metastatic cancer cells are highly plastic for the expression of different tumor phenotype hallmarks and organotropism. This plasticity is highly regulated but the dynamics of the signaling processes orchestrating the shift from one cell phenotype and metastatic organ pattern to another are still largely unknown. The scaffolding protein NHERF1 has been shown to regulate the expression of different neoplastic phenotypes through its PDZ domains, which forms the mechanistic basis for metastatic organotropism. This reprogramming activity was postulated to be dependent on its differential phosphorylation patterns. Here, we show that NHERF1 phosphorylation on S279/S301 dictates several tumor phenotypes such as in vivo invasion, NHE1-mediated matrix digestion, growth and vasculogenic mimicry. Remarkably, injecting mice with cells having differential NHERF1 expression and phosphorylation drove a shift from the predominantly lung colonization (WT NHERF1) to predominately bone colonization (double S279A/S301A mutant), indicating that NHERF1 phosphorylation also acts as a signaling switch in metastatic organotropism.
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- 2019
6. Integrin-Linked Kinase Links Integrin Activation to Invadopodia Function and Invasion via the p(T567)-Ezrin/NHERF1/NHE1 Pathway
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Maria Raffaella Greco, Khalid O. Alfarouk, Stephan J. Reshkin, Loredana Moro, Stefania Forciniti, Stefania Cannone, and Rosa Angela Cardone
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0301 basic medicine ,Scaffold protein ,Male ,Sodium-Hydrogen Exchangers ,Integrin ,Proximity ligation assay ,Protein Serine-Threonine Kinases ,Article ,Catalysis ,lcsh:Chemistry ,Inorganic Chemistry ,Extracellular matrix ,03 medical and health sciences ,breast cancer ,0302 clinical medicine ,Ezrin ,Cell Line, Tumor ,Humans ,Integrin-linked kinase ,Physical and Theoretical Chemistry ,lcsh:QH301-705.5 ,Molecular Biology ,Spectroscopy ,invadopodia ,integrin signaling ,Sodium-Hydrogen Exchanger 1 ,biology ,pH ,Chemistry ,Integrin beta1 ,Organic Chemistry ,General Medicine ,prostate cancer ,invasion ,Phosphoproteins ,Computer Science Applications ,Cell biology ,Extracellular Matrix ,Cytoskeletal Proteins ,030104 developmental biology ,lcsh:Biology (General) ,lcsh:QD1-999 ,030220 oncology & carcinogenesis ,Invadopodia ,PC-3 Cells ,Podosomes ,biology.protein ,Signal transduction - Abstract
Tumor cell invasion depends largely on degradation of the extracellular matrix (ECM) by protease-rich structures called invadopodia, whose formation and activity requires the convergence of signaling pathways engaged in cell adhesion, actin assembly, membrane regulation and ECM proteolysis. It is known that β1-integrin stimulates invadopodia function through an invadopodial p(T567)-ezrin/NHERF1/NHE1 signal complex that regulates NHE1-driven invadopodia proteolytic activity and invasion. However, the link between β1-integrin and this signaling complex is unknown. In this study, in metastatic breast (MDA-MB-231) and prostate (PC-3) cancer cells, we report that integrin-linked kinase (ILK) integrates β1-integrin with this signaling complex to regulate invadopodia activity and invasion. Proximity ligation assay experiments demonstrate that, in invadopodia, ILK associates with β1-integrin, NHE1 and the scaffold proteins p(T567)-ezrin and NHERF1. Activation of β1-integrin increased both invasion and invadopodia activity, which were specifically blocked by inhibition of either NHE1 or ILK. We conclude that ILK integrates β1-integrin with the ECM proteolytic/invasion signal module to induce NHE1-driven invadopodial ECM proteolysis and cell invasion.
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- 2021
7. Rett Syndrome around the world
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Gabriella Martino, Tindara Caprì, and Rosa Angela Fabio
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Pediatrics ,medicine.medical_specialty ,medicine ,Rett syndrome ,Biology ,medicine.disease - Published
- 2019
8. The Possible Role of Helicobacter pylori in Gastric Cancer and Its Management
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Khalid O. Alfarouk, Adil H. H. Bashir, Ahmed N. Aljarbou, AbdelRahman M. Ramadan, Abdel Khalig Muddathir, Sari T. S. AlHoufie, Abdelhamid Hifny, Gamal O. Elhassan, Muntaser E. Ibrahim, Saad S. Alqahtani, Shakir D. AlSharari, Claudiu T. Supuran, Cyril Rauch, Rosa Angela Cardone, Stephan J. Reshkin, Stefano Fais, and Salvador Harguindey
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0301 basic medicine ,Cancer Research ,Anaerobic bacterium ,Review ,lcsh:RC254-282 ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Normal flora ,Carcinogen ,Related factors ,Helicobacter pylori ,biology ,pH ,business.industry ,gastric cancer ,Cancer ,lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,medicine.disease ,biology.organism_classification ,030104 developmental biology ,Oncology ,inflammation ,030220 oncology & carcinogenesis ,Cancer research ,pharmacology ,business - Abstract
Helicobacter pylori (HP) is a facultative anaerobic bacterium. HP is a normal flora having immuno-modulating properties. This bacterium is an example of a microorganism inducing gastric cancer. Its carcinogenicity depends on bacteria-host related factors. The proper understanding of the biology of HP inducing gastric cancer offers the potential strategy in the managing of HP rather than eradicating it. In this article, we try to summarize the biology of HP-induced gastric cancer and discuss the current pharmacological approach to treat and prevent its carcinogenicity.
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- 2019
9. Extracellular matrix degradation via enolase/plasminogen interaction: Evidence for a mechanism conserved in Metazoa
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Rossana Girardello, Annalisa Grimaldi, Elena Coviello, Stephan J. Reshkin, Patrizia Falabella, Gerarda Grossi, Maria Raffaella Greco, Magnus Monné, Simona Laurino, and Rosa Angela Cardone
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0301 basic medicine ,Signal peptide ,Enolase ,Cell Biology ,General Medicine ,Biology ,Embryonic stem cell ,Cell membrane ,Extracellular matrix ,03 medical and health sciences ,030104 developmental biology ,medicine.anatomical_structure ,Biochemistry ,medicine ,Extracellular ,Glycolysis ,Extracellular Matrix Degradation - Abstract
Background Information While enolase is a ubiquitous metalloenzyme involved in the glycolytic pathway, it is also known as a multifunctional protein, since enolases anchored on the outer surface of the plasma membrane are involved in tissue invasion. Results We have identified an extracellular enolase (Ae-ENO) produced by the teratocytes, embryonic cells of the insect parasitoid Aphidius ervi. We demonstrate that Ae-ENO, although lacking a signal peptide, accumulates in cytoplasmic vesicles oriented towards the cell membrane. Ae-ENO binds to and activates a plasminogen-like molecule inducing digestion of the host tissue and thereby ensuring successful parasitism. Conclusions These results support the hypothesis that plasminogen-like proteins exist in invertebrates. Interestingly the activation of a plasminogen-like protein is mediated by a mechanisms involving the surface enolase/fibrinolytic system considered, until now, exclusive of vertebrates, and that instead is conserved across species. Significance To our knowledge, this is the first example of enolase mediated Plg-like binding and activation in insect cells, demonstrating the existence of an ECM degradation process via a Plg-like protein in invertebrates.
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- 2016
10. Extracellular Matrix composition modulates PDAC parenchymal and stem cell plasticity and behavior through the secretome
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Elisa Dalla Pozza, Katrine Zeeberg, Ilaria Dando, Stefania Forciniti, Maria Mastrodonato, Maria Raffaella Greco, Stefania Cannone, Stephan J. Reshkin, Giulia Biondani, Valeria Casavola, Marta Palmieri, and Rosa Angela Cardone
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0301 basic medicine ,Vascular Endothelial Growth Factor A ,Angiogenesis ,Population ,Cell Plasticity ,Cell Culture Techniques ,Biology ,Adenocarcinoma ,Biochemistry ,Collagen Type I ,Extracellular matrix ,03 medical and health sciences ,Cancer stem cell ,Cell Line, Tumor ,Tumor Microenvironment ,Humans ,Vasculogenic mimicry ,Neoplasm Invasiveness ,Pancreatic Adenocarcinoma ,Neoplasm Metastasis ,education ,Molecular Biology ,Parenchymal Tissue ,Cell Proliferation ,education.field_of_study ,Tumor microenvironment ,Matrigel ,Neovascularization, Pathologic ,Cancer stem cells ,Desmoplastic Reaction ,Cell Biology ,Vascular Endothelial Growth Factor Receptor-2 ,3D organotypic cultures ,VEFGR-2 ,Extracellular Matrix ,Gene Expression Regulation, Neoplastic ,030104 developmental biology ,Cancer research ,Neoplastic Stem Cells ,Stem cell ,Carcinoma, Pancreatic Ductal - Abstract
Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal cancers. Its aggressiveness is driven by an intense fibrotic desmoplastic reaction in which the increasingly collagen I-rich extracellular matrix (ECM) and several cell types, including cancer stem cells (CSCs), create a tumor-supportive environment. However, how ECM composition regulates CSC dynamics and their relationship with the principle parenchymal tumor population to promote early invasive growth is not yet characterized. For this, we utilized a platform of 3D organotypic cultures composed of laminin-rich Matrigel, representative of an early tumor, plus increasing concentrations of collagen I to simulate malignant stroma progression. As ECM collagen I increases, CSCs progress from a rapidly growing, vascular phenotype to a slower growing, avascular phase, while maintaining their endothelial-like gene signatures. This transition is supported autocrinically by the CSCs and paracrinically by the parenchymal cells via their ECM-dependent secretomes. Indeed, when growing on an early tumor ECM, the CSCs are dedicated toward the preparation of a vascular niche by (a) activating their growth program, (b) secreting high levels of proangiogenic factors which stimulate both angiogenesis and vasculogenic mimicry, and (c) overexpressing VEGFR-2, which is activated by VEGF secreted by both the CSC and parenchymal cells. On Matrigel, the more differentiated parenchymal tumor cell population had reduced growth but a high invasive capacity. This concerted high local invasion of parenchymal cells into the CSC-derived vascular network suggests that a symbiotic relationship between the parenchymal cells and the CSCs underlies the initiation and maintenance of early PDAC infiltration and metastasis.
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- 2018
11. A Novel NHE1-Centered Signaling Cassette Drives Epidermal Growth Factor Receptor–Dependent Pancreatic Tumor Metastasis and Is a Target for Combination Therapy
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Frauke Alves, Angela Zaccagnino, Antonia Bellizzi, Maria Raffaella Greco, Rosa Angela Cardone, Stephan J. Reshkin, Philipp Bruns, Albrecht Schwab, Valeria Casavola, Holger Kalthoff, Mara Saccomano, Christian Pilarsky, Marta Menga, and Katrine Zeeberg
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Scaffold protein ,Cancer Research ,medicine.disease_cause ,lcsh:RC254-282 ,Metastasis ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Pancreatic tumor ,medicine ,Epidermal growth factor receptor ,030304 developmental biology ,0303 health sciences ,biology ,Cariporide ,medicine.disease ,lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,3. Good health ,chemistry ,Cell culture ,030220 oncology & carcinogenesis ,Immunology ,Cancer research ,biology.protein ,Erlotinib ,KRAS ,medicine.drug - Abstract
Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal cancers principally because of early invasion and metastasis. The epidermal growth factor receptor (EGFR) is essential for PDAC development even in the presence of Kras, but its inhibition with erlotinib gives only a modest clinical response, making the discovery of novel EGFR targets of critical interest. Here, we revealed by mining a human pancreatic gene expression database that the metastasis promoter Na+/H+ exchanger (NHE1) associates with the EGFR in PDAC. In human PDAC cell lines, we confirmed that NHE1 drives both basal and EGF-stimulated three-dimensional growth and early invasion via invadopodial extracellular matrix digestion. EGF promoted the complexing of EGFR with NHE1 via the scaffolding protein Na+/H+ exchanger regulatory factor 1, engaging EGFR in a negative transregulatory loop that controls the extent and duration of EGFR oncogenic signaling and stimulates NHE1. The specificity of NHE1 for growth or invasion depends on the segregation of the transient EGFR/Na+/H+ exchanger regulatory factor 1/NHE1 signaling complex into dimeric subcomplexes in different lipid raftlike membrane domains. This signaling complex was also found in tumors developed in orthotopic mice. Importantly, the specific NHE1 inhibitor cariporide reduced both three-dimensional growth and invasion independently of PDAC subtype and synergistically sensitized these behaviors to low doses of erlotinib.
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- 2015
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12. The scaffolding protein NHERF1 sensitizes EGFR-dependent tumor growth, motility and invadopodia function to gefitinib treatment in breast cancer cells
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Katrine Zeeberg, Stephan J. Reshkin, Rosa Rubino, Stefania Forciniti, Angelo Paradiso, Rosa Angela Cardone, Antonia Bellizzi, and Maria Raffaella Greco
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Cancer Research ,Sodium-Hydrogen Exchangers ,Triple Negative Breast Neoplasms ,Biology ,Gefitinib ,Downregulation and upregulation ,Cell Movement ,Tumor Cells, Cultured ,medicine ,Humans ,Neoplasm Invasiveness ,Pseudopodia ,Epidermal growth factor receptor ,Triple-negative breast cancer ,Cell Proliferation ,Oncogene ,Plakins ,Cell cycle ,Phosphoproteins ,ErbB Receptors ,Protein Transport ,Oncology ,Drug Resistance, Neoplasm ,Invadopodia ,Quinazolines ,Cancer research ,biology.protein ,Female ,Tyrosine kinase ,medicine.drug - Abstract
Triple negative breast cancer (TNBC) patients cannot be treated with endocrine therapy or targeted therapies due to lack of related receptors. These patients overexpress the epidermal growth factor receptor (EGFR), but are resistant to tyrosine kinase inhibitors (TKIs) and anti-EGFR therapies. Mechanisms suggested for resistance to TKIs include EGFR independence, mutations and alterations in EGFR and in its downstream signalling pathways. Ligand-induced endocytosis and degradation of EGFR play important roles in the downregulation of the EGFR signal suggesting that its activity could be regulated by targeting its trafficking. Evidence in normal cells showing that the scaffolding protein Na+/H+ exchanger regulatory factor 1 (NHERF1) can associate with EGFR to regulate its trafficking, led us to hypothesize that NHERF1 expression levels could regulate EGFR trafficking and functional expression in TNBC cells and, in this way, modulate its role in progression and response to treatment. We investigated the subcellular localization of NHERF1 and its interaction with EGFR in a metastatic basal like TNBC cell model, MDA-MB‑231, and the role of forced NHERF1 overexpression and/or stimulation with EGF on the sensitivity to EGFR specific TKI treatment with gefitinib. Stimulation with EGF induces an interaction of NHERF1 with EGFR to regulate its localization, degradation and function. NHERF1 overexpression is sufficient to drive its interaction with EGFR in non-stimulated conditions, inhibits EGFR degradation and increases its retention time in the plasma membrane. Importantly, NHERF1 overexpression strongly sensitized the cell to the pharmacological inhibition by gefitinib of EGFR-driven growth, motility and invadopodia-dependent ECM proteolysis. The further determination of how the NHERF1‑EGFR interaction is regulated may improve our understanding of TNBC resistance to the action of existing anticancer drugs.
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- 2014
13. Glycolysis, tumor metabolism, cancer growth and dissemination. A new pH-based etiopathogenic perspective and therapeutic approach to an old cancer question
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H H Bashir Adil, Cyril Rauch, Abdel Khalig Muddathir, Muntaser E. Ibrahim, Gamal O. Elhassan, Khalid O. Alfarouk, Julián Polo Orozco, Stephan J. Reshkin, Salvador Harguindey, Daniel Verduzco, and Rosa Angela Cardone
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Cancer Research ,Biology ,Bioinformatics ,cancer growth ,cancer treatment ,Therapeutic approach ,medicine ,pH and cancer ,metastatic process ,Glycolysis ,pH and glycolysis ,Tumor glycolysis ,Cancer ,new paradigm in oncology ,Transporter ,Metabolism ,proton transport inhibitors ,Hypoxia (medical) ,medicine.disease ,Warburg effect ,Oncology ,Research Perspective ,Cancer cell ,Cancer research ,Erratum ,medicine.symptom - Abstract
Cancer cells acquire an unusual glycolytic behavior relative, to a large extent, to their intracellular alkaline pH (pHi). This effect is part of the metabolic alterations found in most, if not all, cancer cells to deal with unfavorable conditions, mainly hypoxia and low nutrient supply, in order to preserve its evolutionary trajectory with the production of lactate after ten steps of glycolysis. Thus, cancer cells reprogram their cellular metabolism in a way that gives them their evolutionary and thermodynamic advantage. Tumors exist within a highly heterogeneous microenvironment and cancer cells survive within any of the different habitats that lie within tumors thanks to the overexpression of different membrane-bound proton transporters. This creates a highly abnormal and selective proton reversal in cancer cells and tissues that is involved in local cancer growth and in the metastatic process. Because of this environmental heterogeneity, cancer cells within one part of the tumor may have a different genotype and phenotype than within another part. This phenomenon has frustrated the potential of single-target therapy of this type of reductionist therapeutic approach over the last decades. Here, we present a detailed biochemical framework on every step of tumor glycolysis and then proposea new paradigm and therapeutic strategy based upon the dynamics of the hydrogen ion in cancer cells and tissues in order to overcome the old paradigm of one enzyme-one target approach to cancer treatment. Finally, a new and integral explanation of the Warburg effect is advanced.
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- 2014
14. Cellular acidification as a new approach to cancer treatment and to the understanding and therapeutics of neurodegenerative diseases
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Salvador Harguindey, Gorka Orive, Eduardo Anitua, Rosa Angela Cardone, Khalid O. Alfarouk, Jesús Devesa, Stephan J. Reshkin, Cyril Rauch, Sébastien Roger, Julián Polo Orozco, Pablo Devesa, and Daniel Stanciu
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0301 basic medicine ,Cancer Research ,Neurodegeneration ,Cancer ,Apoptosis ,Neurodegenerative Diseases ,Disease ,Biology ,Hydrogen-Ion Concentration ,medicine.disease ,Cell biology ,03 medical and health sciences ,030104 developmental biology ,Mediator ,Proton transport ,Cancer cell ,medicine ,Cancer research ,Humans ,Acids ,Intracellular ,Homeostasis - Abstract
During the last few years, the understanding of the dysregulated hydrogen ion dynamics and reversed proton gradient of cancer cells has resulted in a new and integral pH-centric paradigm in oncology, a translational model embracing from cancer etiopathogenesis to treatment. The abnormalities of intracellular alkalinization along with extracellular acidification of all types of solid tumors and leukemic cells have never been described in any other disease and now appear to be a specific hallmark of malignancy. As a consequence of this intracellular acid-base homeostatic failure, the attempt to induce cellular acidification using proton transport inhibitors and other intracellular acidifiers of different origins is becoming a new therapeutic concept and selective target of cancer treatment, both as a metabolic mediator of apoptosis and in the overcoming of multiple drug resistance (MDR). Importantly, there is increasing data showing that different ion channels contribute to mediate significant aspects of cancer pH regulation and etiopathogenesis. Finally, we discuss the extension of this new pH-centric oncological paradigm into the opposite metabolic and homeostatic acid-base situation found in human neurodegenerative diseases (HNDDs), which opens novel concepts in the prevention and treatment of HNDDs through the utilization of a cohort of neural and non-neural derived hormones and human growth factors.
- Published
- 2016
15. Protease activity at invadopodial focal digestive areas is dependent on NHE1-driven acidic pHe
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Rosa Rubino, Lorenzo Guerra, Ester Antelmi, Maria Raffaella Greco, Stephan J. Reshkin, Valeria Casavola, Giovanni Busco, Rosa Angela Cardone, Centre de biophysique moléculaire (CBM), Université d'Orléans (UO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), Dept Biosci Biotechnol & Bropharmaceut, Università degli Studi di Bari Aldo Moro, Department of General and Environmental Physiology, and Università degli studi di Bari Aldo Moro (UNIBA)
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Cancer Research ,Proteases ,Sodium-Hydrogen Exchangers ,Phenylalanine ,medicine.medical_treatment ,Proteolysis ,Breast Neoplasms ,[SDV.CAN]Life Sciences [q-bio]/Cancer ,Thiophenes ,[SDV.BC]Life Sciences [q-bio]/Cellular Biology ,Matrix Metalloproteinase Inhibitors ,Biology ,Guanidines ,Cathepsin B ,Extracellular matrix ,Serine ,03 medical and health sciences ,0302 clinical medicine ,Cell Line, Tumor ,Matrix Metalloproteinase 14 ,Extracellular ,medicine ,Humans ,Neoplasm Invasiveness ,Secretion ,Sulfones ,[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM] ,Cation Transport Proteins ,ComputingMilieux_MISCELLANEOUS ,030304 developmental biology ,0303 health sciences ,Sodium-Hydrogen Exchanger 1 ,Protease ,medicine.diagnostic_test ,[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology ,General Medicine ,Hydrogen-Ion Concentration ,Extracellular Matrix ,Cell biology ,Matrix Metalloproteinase 9 ,Oncology ,Biochemistry ,030220 oncology & carcinogenesis ,Invadopodia ,Matrix Metalloproteinase 2 ,Female ,Cell Surface Extensions ,Anti-Arrhythmia Agents ,Peptide Hydrolases - Abstract
Degradation of the extracellular matrix (ECM) is a critical step of tumor cell invasion and requires protease- dependent proteolysis focalized at the invadopodia where the proteolysis of the ECM occurs. Most of the extracellular proteases belong to serine- or metallo-proteases and the inva- dopodia is where protease activity is regulated. While recent data looking at global protease activity in the growth medium reported that their activity and role in invasion is dependent on Na + /H + exchanger 1 (NHE1)-driven extracellular acidifica - tion, there is no data on this aspect at the invadopodia, and an open question remains whether this acid extracellular pH (pHe) activation of proteases in tumor cells occurs pref- erentially at invadopodia. We previously reported that the NHE1 is expressed in breast cancer invadopodia and that the NHE1-dependent acidification of the peri-invadopodial space is critical for ECM proteolysis. In the present study, using, for the first time, in situ zymography analysis, we demonstrated a concordance between NHE1 activity, extracellular acidifi - cation and protease activity at invadopodia to finely regulate ECM digestion. We demonstrated that: (i) ECM proteolysis taking place at invadopodia is driven by acidification of the peri-invadopodia microenvironment; (ii) that the proteases have a functional pHe optimum that is acidic; (iii) more than one protease is functioning to digest the ECM at these invadopodial sites of ECM proteolysis; and (iv) lowering pHe or inhibiting the NHE1 increases protease secretion while blocking protease activity changes NHE1 expression at the invadopodia.
- Published
- 2013
16. MED1101: A new dialdehydic compound regulating P2×7 receptor cell surface expression in U937 cells
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Stephan J. Reshkin, Valeria Casavola, Domenico Marzulli, Elena Ciani, Stefania Muzzachi, Giulia Merizzi, Rosa Angela Cardone, Antonella Blasi, Maria Favia, Lorenzo Guerra, and Antonio Soleti
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U937 cell ,Microglia ,Monocyte ,Purinergic receptor ,Inflammation ,Cell Biology ,General Medicine ,Biology ,Cell biology ,Intracellular signal transduction ,medicine.anatomical_structure ,medicine ,medicine.symptom ,Signal transduction ,Receptor - Abstract
Background information P2×7R is a member of the ionotropic family of purinergic receptors activated by millimolar concentrations of extracellular ATP such as induced by inflammatory stimuli. The receptor is widely expressed in cells of haematopoietic origin such as monocytes, macrophages and microglia. There is growing interest in anta-gonist compounds of the P2×7R since it has been demonstrated to be a viable therapeutic target for inflammatory diseases. Here, we tested the possible P2×7 antagonist effect of MED1101, a newly synthesised dialdehydic compound on U937 monocyte cells. Results Human U937 cells express the full-length P2×7A receptor isoform. Treatment with lipopolysaccharide (LPS), a potent inducer of inflammation, significantly increased the expression of the receptor in the plasma membrane. Importantly, MED1101 induced internalisation of the P2×7R already after 30 min incubation in both physiological conditions and in presence of the inflammatory stimulus (LPS) and this effect was observable for up to 12 h after its removal. Moreover, MED1101 induced an impairment of monocyte migration/transmigration through direct P2×7R antagonism and subsequent inhibition of the intracellular signal transduction processes of Ca2+ influx and MAPK phosphorylation. Conclusions Our results clearly demonstrate that in U937 monocyte cells MED1101 acts as a P2×7R antagonist through the induction of receptor internalisation and subsequent inhibition of down-stream signal transduction pathways that regulate monocyte migration/transmigration, thus playing a potential therapeutic role in inflammatory diseases.
- Published
- 2013
17. CLA Reduces Inflammatory Mediators from A427 Human Lung Cancer Cells and A427 Conditioned Medium Promotes Differentiation of C2C12 Murine Muscle Cells
- Author
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Manuela Oraldi, Elena Paiuzzi, Giuliana Muzio, Rosa Angela Canuto, and Marina Maggiora
- Subjects
Indoles ,Lung Neoplasms ,Cachexia ,medicine.medical_treatment ,Cellular differentiation ,Peroxisome Proliferator-Activated Receptors ,Muscle cells ,Human lung cancer ,Peroxisome proliferator-activated receptor ,Biology ,Biochemistry ,Dinoprostone ,Mice ,Cell Line, Tumor ,medicine ,Animals ,Anticarcinogenic Agents ,Myocyte ,Linoleic Acids, Conjugated ,Horses ,Interleukin 6 ,Lung ,Cell Proliferation ,chemistry.chemical_classification ,integumentary system ,Interleukin-6 ,Conjugated linoleic acid ,Cytokines ,Organic Chemistry ,food and beverages ,Interleukin ,Cell Differentiation ,Cell Biology ,Cytokine ,chemistry ,Cell culture ,Culture Media, Conditioned ,Immunology ,biology.protein ,Cancer research ,lipids (amino acids, peptides, and proteins) ,Tumor necrosis factor alpha ,Inflammation Mediators ,Interleukin-1 - Abstract
Conjugated linoleic acid (CLA) is thought to have anti-proliferative and anti-inflammatory properties, but its effect on cancer cachexia is unknown. Two effects were here investigated: that of CLA on inflammatory mediator production in human lung cancer cells, and that of reduced mediators on the myogenic differentiation of murine muscle C2C12 cells. The latter cells were grown in medium conditioned by human lung cancer A427 cells, with or without CLA, to mimic only the effect of molecules released from the tumor "in vivo", excluding the effect of host-produced cachectic factors. The results obtained show that CLA was found to reduce the production of tumor necrosis factor-α, interleukin (IL)-1β and prostaglandin E2 (PGE2), but had no effect on IL-6 production. The mechanisms underlying the effect of CLA on cytokine or PGE2 release in A427 cells are probably mediated by activation of peroxisome proliferator-activated receptor (PPAR)α, which increased at 24 h CLA treatment. In turn, the reduced content of inflammatory mediators in medium conditioned by A427 cells, in the presence of CLA, allowed muscle cells to proliferate, again by inducing PPAR. The involvement of PPARα was demonstrated by treatment with the antagonist MK-886. The findings demonstrate the anti-inflammatory and myogenic action of CLA and point to its possible application as a novel dietary supplement and therapeutic agent in inflammatory disease states, such as cachexia.
- Published
- 2012
18. Assessment of different 3D culture systems to study tumor phenotype and chemosensitivity in pancreatic ductal adenocarcinoma
- Author
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Maria Raffaella Greco, Katrine Zeeberg, Stephan J. Reshkin, Mara Saccomano, Stine F. Pedersen, Frauke Alves, Rosa Angela Cardone, and Asbjørn Nøhr-Nielsen
- Subjects
0301 basic medicine ,Cancer Research ,Pathology ,medicine.medical_specialty ,Cell Culture Techniques ,Biology ,Adenocarcinoma ,03 medical and health sciences ,Erlotinib Hydrochloride ,Mice ,In vivo ,Cell Line, Tumor ,medicine ,Animals ,Humans ,Epidermal growth factor receptor ,Cell Proliferation ,Matrigel ,Oncogene ,Epidermal Growth Factor ,Cell cycle ,Molecular medicine ,ErbB Receptors ,030104 developmental biology ,Oncology ,Cell culture ,Drug Resistance, Neoplasm ,Cancer research ,biology.protein ,Erlotinib ,Stromal Cells ,medicine.drug ,Carcinoma, Pancreatic Ductal - Abstract
Pancreatic ductal adenocarcinoma (PDAC) is a highly malignant disease with a very poor prognosis, due to the influence of the tumor stroma, which promotes tumor growth, early invasion and chemoradiation resistance. Efforts to develop models for identifying novel anticancer therapeutic compounds have been hampered by the limited ability of in vitro models to mimic these in vivo tumor-stroma interactions. This has led to the development of various three-dimensional (3D) culture platforms recapitulating the in vivo tumor-stroma crosstalk and designed to better understand basic cancer processes and screen drug action. However, a consensus for different experimental 3D platforms is still missing in PDAC. We compared four PDAC cell lines of different malignancy grown in 2D monolayers to three of the more commonly used 3D techniques (ultralow adhesion concave microwells, Matrigel inclusion and organotypic systems) and to tumors derived from their orthotopic implantation in mice. In these 3D platforms, we observed that cells grow with very different tumor morphologies and the organotypic setting most closely resembles the tumor cytoarchitecture obtained by orthotopically implanting the four cell lines in mice. We then analyzed the molecular and cellular responses of one of these cell lines to epidermal growth factor receptor (EGFR) stimulation with EGF and inhibition with erlotinib and found that only in the 3D platforms, and especially the organotypic, cells: i) responded to EGF by changing the expression of signalling components underlying cell-stroma crosstalk and tissue architecture, growth, invasion and drug resistance (E-cadherin, EGFR, ezrin, β1 integrin, NHERF1 and HIF-1α) similar to those reported in vivo; ii) had stimulated growth and increased erlotinib sensitivity in response to EGF, more faithfully mimicking their known in vivo behaviour. Altogether, these results, indicate the organotypic as the most relevant physiological 3D system to study the complex tumor stroma interactions driving progression and determining chemio-resistance.
- Published
- 2016
19. Correctors of mutant CFTR enhance subcortical cAMP/PKA signaling via ezrin phosphorylation and cytoskeleton organization
- Author
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Manuela Zaccolo, Maria Teresa Mancini, Valeria Casavola, Onofrio Laselva, Lorenzo Guerra, Stefano Castellani, Massimo Conese, Anna Claudia Abbattiscianni, Stefania Monterisi, Maria Favia, Francesca Di Sole, and Rosa Angela Cardone
- Subjects
0301 basic medicine ,Cytoskeleton organization ,macromolecular substances ,Cell Biology ,Biology ,Apical membrane ,Actin cytoskeleton ,Cystic fibrosis transmembrane conductance regulator ,Cell biology ,03 medical and health sciences ,030104 developmental biology ,Ezrin ,Mutant protein ,biology.protein ,Phosphorylation ,Cytoskeleton - Abstract
The most common mutation of the cystic fibrosis transmembrane regulator (CFTR) gene, F508del, produces a misfolded protein resulting in its defective trafficking to the cell surface and an impaired chloride secretion. Pharmacological treatments partially rescue F508del CFTR activity either directly by interacting with the mutant protein and/or indirectly by altering the cellular protein homeostasis. Here, we show that the phosphorylation of ezrin together with its binding to phosphatidylinositol-4,5-bisphosphate (PIP2) tethers the F508del CFTR to the actin cytoskeleton, stabilizing it on the apical membrane and rescuing the sub-membrane compartmentalization of cAMP and activated PKA. Both the small molecules trimethylangelicin (TMA) and VX-809, which act as 'correctors' for F508del CFTR by rescuing F508del-CFTR-dependent chloride secretion, also restore the apical expression of phosphorylated ezrin and actin organization and increase cAMP and activated PKA submembrane compartmentalization in both primary and secondary cystic fibrosis airway cells. Latrunculin B treatment or expression of the inactive ezrin mutant T567A reverse the TMA and VX-809-induced effects highlighting the role of corrector-dependent ezrin activation and actin re-organization in creating the conditions to generate a sub-cortical cAMP pool of adequate amplitude to activate the F508del-CFTR-dependent chloride secretion.
- Published
- 2016
20. 4-Hydroxyhexenal and 4-hydroxynonenal are mediators of the anti-cachectic effect of n-3 and n-6 polyunsaturated fatty acids on human lung cancer cells
- Author
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Marina Ricci, Fiammetta Monacelli, Giuliana Muzio, Manuela Oraldi, Nicola Traverso, Marina Maggiora, and Rosa Angela Canuto
- Subjects
0301 basic medicine ,Cachexia ,Cellular differentiation ,Muscle cells ,PPRE ,Biochemistry ,Lipid peroxidation ,chemistry.chemical_compound ,Mice ,0302 clinical medicine ,Myosin ,TNFα ,Myocyte ,Cell Differentiation ,lipid peroxidation ,Eicosapentaenoic Acid ,030220 oncology & carcinogenesis ,HHE ,lipids (amino acids, peptides, and proteins) ,Tumor necrosis factor alpha ,Lung cancer ,Protein Binding ,medicine.medical_specialty ,HNE ,n-3 PUFAs ,n-6 PUFAs, lipid peroxidation ,PPARs ,Physiology (medical) ,Docosahexaenoic Acids ,Respiratory Mucosa ,Biology ,4-Hydroxynonenal ,Cell Line ,03 medical and health sciences ,Internal medicine ,Cell Line, Tumor ,medicine ,Animals ,Humans ,PPAR alpha ,Cell Proliferation ,Aldehydes ,Binding Sites ,Myosin Heavy Chains ,Cell growth ,Tumor Necrosis Factor-alpha ,n-6 PUFAs ,030104 developmental biology ,Endocrinology ,chemistry ,Gene Expression Regulation ,Cell culture ,Culture Media, Conditioned - Abstract
Cachexia, the most severe paraneoplastic syndrome, occurs in about 80% of patients with advanced cancer; it cannot be reverted by conventional, enteral, or parenteral nutrition. For this reason, nutritional interventions must be based on the use of substances possessing, alongside nutritional and energetic properties, the ability to modulate production of the pro-inflammatory factors responsible for the metabolic changes characterising cancer cachexia. In light of their nutritional and anti-inflammatory properties, polyunsaturated fatty acids (PUFAs), and in particular n-3, have been investigated for treating cachexia; however, the results have been contradictory. Since both n-3 and n-6 PUFAs can affect cell functions in several ways, this research investigated the possibility that the effects of both n-3 and n-6 PUFAs could be mediated by their major aldehydic products of lipid peroxidation, 4-hydroxyhexenal (HHE) and 4-hydroxynonenal (HNE), and by their anti-inflammatory properties. An "in vitro" cancer cachexia model, consisting of human lung cancer cells (A427) and murine myoblasts (C2C12), was used. The results showed that: 1) both n-3 and n-6 PUFAs reduced the growth of lung cancer cells without causing cell death, increased lipid peroxidation and Peroxisome Proliferator-Activated Receptor (PPAR)α, and decreased TNFα; 2) culture medium conditioned by A427 cells grown in the absence of PUFAs blocked myosin production and the differentiation of C2C12 muscle cells; conversely, muscle cells grown in culture medium conditioned by the same cells in the presence of PUFAs showed myosin expression and formed myotubes; 3) adding HHE or HNE directly to C2C12 cells maintained in culture medium conditioned by A427 cells in the absence of PUFAs stimulated myosin production and myotube formation; 4) putative consensus sequences for (PPARs) have been found in genes encoding fast isoforms of myosin heavy chain, by a bioinformatics approach. The overall results show, first, the ability of both n-3 and n-6 PUFAs and their lipid peroxidation products to prevent the blocking of myosin expression and myotube formation caused in C2C12 cells by medium conditioned by human lung tumour cells. The C2C12 cell differentiation can be due to direct effect of lipid peroxidation products, as evidenced by treating C2C12 cells with HHE and HNE, and to the decrease of pro-inflammatory TNFα in A427 cell culture medium. The presence of consensus sequences for PPARs in genes encoding the fast isoforms of myosin heavy chain suggests that the effects of PUFAs, HHE, and HNE are PPAR-mediated.
- Published
- 2016
21. Molecular pathways shared between host-parasitoid interaction in insect and other animals: The case of teratocyte extracellular enolase
- Author
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Gerarda Grossi, Simona Laurino, Annalisa Grimaldi, Rosa Angela Cardone, and Patrizia Falabella
- Subjects
biology ,Host (biology) ,media_common.quotation_subject ,Enolase ,Insect ,biology.organism_classification ,teratocytes ,Hymenoptera, teratocytes, enolase, plasminogen, extracellular matrix digestion ,Hymenoptera ,Parasitoid ,enolase ,extracellular matrix digestion ,Biochemistry ,Extracellular ,plasminogen ,media_common - Published
- 2016
22. NHERF1 acts as a molecular switch to program metastatic behavior and organotropism via its PDZ domains
- Author
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Cardone, Rosa Angela, GRECO, MARIA RAFFAELLA, Capulli, Mattia, Weinman, Edward, Busco, Giovanni, Bellizzi, Antonia, Casavola, Valeria, Antelmi, Ester, Ambruosi, Barbara, Dell'Aquila, Maria Elena, Paradiso, Giovanni, Teti, Anna, Rucci, Nadia, Reshkin, Stephan Joel, Yap, Alpha, Department of General and Environmental Physiology, Università degli studi di Bari, Centre de biophysique moléculaire (CBM), Université d'Orléans (UO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Clinical Experimental Oncology Laboratory, National Cancer Centre, Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Centre National de la Recherche Scientifique (CNRS)-Université de Tours-Institut Français du Cheval et de l'Equitation [Saumur]-Institut National de la Recherche Agronomique (INRA), Section of Veterinary Clinics and Animal Productions, Department Emergency and Organ Transplantation (DETO), University of Bari Aldo Moro (UNIBA), Regenerative Medicine Unit, Ospedale Pediatrico Bambino Gesu` , Rome, Italy, Università degli studi di Bari Aldo Moro (UNIBA), Université d'Orléans (UO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS), Laboratoire Image, Ville, Environnement (LIVE), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), and Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)
- Subjects
Sodium-Hydrogen Exchangers ,Podosome ,PDZ domain ,PDZ Domains ,Apoptosis ,[SDV.CAN]Life Sciences [q-bio]/Cancer ,Biology ,Models, Biological ,Tropism ,Mice ,Structure-Activity Relationship ,03 medical and health sciences ,0302 clinical medicine ,Cell Line, Tumor ,Cell Adhesion ,Human Umbilical Vein Endothelial Cells ,Animals ,Humans ,Neoplasm Invasiveness ,Vasculogenic mimicry ,Pseudopodia ,Neoplasm Metastasis ,[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM] ,Cell adhesion ,Cytoskeleton ,Molecular Biology ,ComputingMilieux_MISCELLANEOUS ,Cell Proliferation ,030304 developmental biology ,Mice, Inbred BALB C ,0303 health sciences ,Neovascularization, Pathologic ,Cell growth ,[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology ,Articles ,Cell Biology ,Transfection ,Phosphoproteins ,Xenograft Model Antitumor Assays ,Phenotype ,Extracellular Matrix ,[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM] ,3. Good health ,Cell biology ,Cell Biology of Disease ,030220 oncology & carcinogenesis ,Female - Abstract
Tumor metastasis is the primary cause of death in cancer patients, but the molecular mechanisms driving the evolution of the phenotype toward a specific organ is one of its less understood aspects. The scaffolding protein NHERF1 reprograms the metastatic phenotype and organotropism via the differential function of its PDZ domains., Metastatic cells are highly plastic for differential expression of tumor phenotype hallmarks and metastatic organotropism. The signaling proteins orchestrating the shift of one cell phenotype and organ pattern to another are little known. Na+/H+ exchanger regulatory factor (NHERF1) is a molecular pathway organizer, PDZ-domain protein that recruits membrane, cytoplasmic, and cytoskeletal signaling proteins into functional complexes. To gain insight into the role of NHERF1 in metastatic progression, we stably transfected a metastatic breast cell line, MDA-MB-231, with an empty vector, with wild-type NHERF1, or with NHERF1 mutated in either the PDZ1- or PDZ2-binding domains to block their binding activities. We observed that NHERF1 differentially regulates the expression of two phenotypic programs through its PDZ domains, and these programs form the mechanistic basis for metastatic organotropism. The PDZ2 domain promotes visceral metastases via increased invadopodia-dependent invasion and anchorage-independent growth, as well as by inhibition of apoptosis, whereas the PDZ1 domain promotes bone metastases by stimulating podosome nucleation, motility, neoangiogenesis, vasculogenic mimicry, and osteoclastogenesis in the absence of increased growth or invasion. Collectively, these findings identify NHERF1 as an important signaling nexus for coordinating cell structure with metastatic behavior and identifies the “mesenchymal-to-vasculogenic” phenotypic transition as an essential step in metastatic progression.
- Published
- 2012
23. Mitochondrial Dysfunction in Cancer and Neurodegenerative Diseases: Spotlight on Fatty Acid Oxidation and Lipoperoxidation Products
- Author
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Fabrizio Gentile, Martina Daga, Alessio Lepore, Giuliana Muzio, Rosa Angela Canuto, Alessia Arcaro, Chiara Dianzani, Giovanni Paolo Cetrangolo, Giuseppina Barrera, Stefania Pizzimenti, and Carlo Ferretti
- Subjects
0301 basic medicine ,Antioxidant ,Physiology ,medicine.medical_treatment ,Clinical Biochemistry ,Oxidative phosphorylation ,Review ,Biology ,Mitochondrion ,Biochemistry ,Lipid peroxidation ,lipoperoxidation products ,03 medical and health sciences ,chemistry.chemical_compound ,mitochondria ,cancer ,neurodegenerative diseases ,fatty acid oxidation ,medicine ,Cardiolipin ,Molecular Biology ,Beta oxidation ,chemistry.chemical_classification ,Reactive oxygen species ,lcsh:RM1-950 ,Cell Biology ,Malondialdehyde ,Cell biology ,lcsh:Therapeutics. Pharmacology ,030104 developmental biology ,chemistry - Abstract
In several human diseases, such as cancer and neurodegenerative diseases, the levels of reactive oxygen species (ROS), produced mainly by mitochondrial oxidative phosphorylation, is increased. In cancer cells, the increase of ROS production has been associated with mtDNA mutations that, in turn, seem to be functional in the alterations of the bioenergetics and the biosynthetic state of cancer cells. Moreover, ROS overproduction can enhance the peroxidation of fatty acids in mitochondrial membranes. In particular, the peroxidation of mitochondrial phospholipid cardiolipin leads to the formation of reactive aldehydes, such as 4-hydroxynonenal (HNE) and malondialdehyde (MDA), which are able to react with proteins and DNA. Covalent modifications of mitochondrial proteins by the products of lipid peroxidation (LPO) in the course of oxidative cell stress are involved in the mitochondrial dysfunctions observed in cancer and neurodegenerative diseases. Such modifications appear to affect negatively mitochondrial integrity and function, in particular energy metabolism, adenosine triphosphate (ATP) production, antioxidant defenses and stress responses. In neurodegenerative diseases, indirect confirmation for the pathogenetic relevance of LPO-dependent modifications of mitochondrial proteins comes from the disease phenotypes associated with their genetic alterations.
- Published
- 2015
24. Superpulsed laser irradiation increases osteoblast activity via modulation of bone morphogenetic factors
- Author
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Germana Martinasso, Renato Pol, Marco Mozzati, Giuliana Muzio, S. Saracino, and Rosa Angela Canuto
- Subjects
Bone sialoprotein ,biology ,business.industry ,Chemistry ,Regeneration (biology) ,Dentistry ,Osteoblast ,Dermatology ,Laser ,law.invention ,medicine.anatomical_structure ,law ,Osteoclast ,Biophysics ,biology.protein ,medicine ,Alkaline phosphatase ,Surgery ,Irradiation ,Osteopontin ,business - Abstract
Background and Objective: Laser therapy is a new approach applicable in different medical fields when bone loss occurs, including orthopedics and dentistry. It has also been used to induce soft-tissue healing, for pain relief, bone, and nerve regeneration. With regard to bone synthesis, laser exposure has been shown to increase osteoblast activity and decrease osteoclast number, by inducing alkaline phosphatase (ALP), osteopontin, and bone sialoprotein expression. Studies have investigated the effects of continuous or pulsed laser irradiation, but no data are yet available on the properties of superpulsed laser irradiation. This study thus aimed to investigate the effect of superpulsed laser irradiation on osteogenic activity of human osteoblast-like cells, paying particular attention to investigating the molecular mechanisms underlying the effects of this type of laser radiation.
- Published
- 2009
25. β-Oestradiol rescues ΔF508CFTR functional expression in human cystic fibrosis airway CFBE41o− cells through the up-regulation of NHERF1
- Author
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Stefania Maria Riccardi, Stephan J. Reshkin, Rosa Angela Cardone, Valeria Casavola, Manuela Zaccolo, Maria Favia, Teresa De Santis, Stefania Monterisi, Teresa Fanelli, and Lorenzo Guerra
- Subjects
Small interfering RNA ,Sodium-Hydrogen Exchangers ,Cystic Fibrosis ,Cell ,Cystic Fibrosis Transmembrane Conductance Regulator ,Gene Expression ,Respiratory Mucosa ,Biology ,Chlorides ,medicine ,Humans ,RNA, Small Interfering ,ΔF508 ,Protein kinase A ,Receptor ,Cells, Cultured ,Estradiol ,Biological Transport ,Cell Biology ,General Medicine ,Transfection ,Apical membrane ,Phosphoproteins ,Cyclic AMP-Dependent Protein Kinases ,Transmembrane protein ,Up-Regulation ,Cell biology ,medicine.anatomical_structure ,Mutation - Abstract
BACKGROUND INFORMATION: CF (cystic fibrosis) is a disease caused by mutations within the CFTR (CF transmembrane conductance regulator) gene. The most common mutation, DeltaF508 (deletion of Phe-508), results in a protein that is defective in folding and trafficking to the cell surface but is functional if properly localized in the plasma membrane. We have recently demonstrated that overexpression of the PDZ protein NHERF1 (Na(+)/H(+)-exchanger regulatory factor 1) in CF airway cells induced both a redistribution of DeltaF508CFTR from the cytoplasm to the apical membrane and the PKA (protein kinase A)-dependent activation of DeltaF508CFTR-dependent chloride secretion. In view of the potential importance of the targeted up-regulation of NHERF1 in a therapeutic context, and since it has been demonstrated that oestrogen treatment increases endogenous NHERF1 expression, we tested the hypothesis that oestrogen treatment can increase NHERF1 expression in a human bronchiolar epithelial CF cell line, CFBE41o(-), with subsequent rescue of apical DeltaF508CFTR chloride transport activity. RESULTS: We found that CFBE41o(-) cells do express ERs (oestrogen receptors) in the nuclear fraction and that beta-oestradiol treatment was able to significantly rescue DeltaF508CFTR-dependent chloride secretion in CFBE41o(-) cell monolayers with a peak between 6 and 12 h of treatment, demonstrating that the DeltaF508CFTR translocated to the apical membrane can function as a cAMP-responsive channel, with a significant increase in chloride secretion noted at 1 nM beta-oestradiol and a maximal effect observed at 10 nM. Importantly, knock-down of NHERF1 expression by transfection with siRNA (small interfering RNA) for NHERF1 inhibited the beta-oestradiol-dependent increase in DeltaF508CFTR protein expression levels and completely prevented the beta-oestradiol-dependent rescue of DeltaF508CFTR transport activity. CONCLUSIONS: These results demonstrate that beta-oestradiol-dependent up-regulation of NHERF1 significantly increases DeltaF508CFTR functional expression in CFBE41o(-) cells.
- Published
- 2008
26. The Omega-3 Fatty Acid Docosahexaenoic Acid Modulates Inflammatory Mediator Release in Human Alveolar Cells Exposed to Bronchoalveolar Lavage Fluid of ARDS Patients
- Author
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Marina Maggiora, Rosa Angela Canuto, Giuliana Muzio, Antonella Trombetta, and Paolo Cotogni
- Subjects
Genetics and Molecular Biology (all) ,ARDS ,Immunology and Microbiology (all) ,lcsh:Medicine ,Biochemistry ,chemistry.chemical_compound ,Omega-6 ,Omega-3 ,Respiratory Distress Syndrome ,Arachidonic Acid ,medicine.diagnostic_test ,Fatty Acids ,NF-kappa B ,General Medicine ,Interleukin-10 ,medicine.anatomical_structure ,Docosahexaenoic acid ,Arachidonic acid ,Inflammation Mediators ,Bronchoalveolar Lavage Fluid ,Cyclooxygenase 2 ,Dinoprostone ,Docosahexaenoic Acids ,Fatty Acids, Omega-3 ,Fatty Acids, Omega-6 ,Gene Expression Regulation ,Humans ,Inflammation ,PPAR gamma ,Pulmonary Alveoli ,Respiratory Distress Syndrome, Adult ,Biochemistry, Genetics and Molecular Biology (all) ,Research Article ,Adult ,medicine.medical_specialty ,Article Subject ,Biology ,General Biochemistry, Genetics and Molecular Biology ,Proinflammatory cytokine ,Alveolar cells ,Internal medicine ,medicine ,Omega 3 fatty acid ,A549 cell ,General Immunology and Microbiology ,lcsh:R ,medicine.disease ,Endocrinology ,Bronchoalveolar lavage ,chemistry ,Immunology - Abstract
Background. This study investigated whether the 1 : 2 ω-3/ω-6 ratio may reduce proinflammatory response in human alveolar cells (A549) exposed to anex vivoinflammatory stimulus (bronchoalveolar lavage fluid (BALF) of acute respiratory distress syndrome (ARDS) patients).Methods. We exposed A549 cells to the BALF collected from 12 ARDS patients. After 18 hours, fatty acids (FA) were added as docosahexaenoic acid (DHA,ω-3) and arachidonic acid (AA,ω-6) in two ratios (1 : 2 or 1 : 7). 24 hours later, in culture supernatants were evaluated cytokines (TNF-α, IL-6, IL-8, and IL-10) and prostaglandins (PGE2and PGE3) release. The FA percentage content in A549 membrane phospholipids, content of COX-2, level of PPARγ, and NF-κB binding activity were determined.Results. The 1 : 2 DHA/AA ratio reversed the baseline predominance ofω-6 overω-3 in the cell membranes (P< 0.001). The proinflammatory cytokine release was reduced by the 1 : 2 ratio (P< 0.01 to P< 0.01). The 1 : 2 ratio reduced COX-2 and PGE2(P< 0.001) as well as NF-κB translocation into the nucleus (P< 0.01), while it increased activation of PPARγand IL-10 release (P< 0.001).Conclusion. This study demonstrated that shifting the FA supply fromω-6 toω-3 decreased proinflammatory mediator release in human alveolar cells exposed to BALF of ARDS patients.
- Published
- 2015
27. Extracellular signal-regulated kinase 1/2 and protein phosphatase 2A are involved in the antiproliferative activity of conjugated linoleic acid in MCF-7 cells
- Author
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Claudia Bocca, Antonella Miglietta, Ludovica Gabriel, Francesca Bozzo, and Rosa Angela Canuto
- Subjects
Cell Survival ,Conjugated linoleic acid ,Medicine (miscellaneous) ,Antineoplastic Agents ,Breast Neoplasms ,Biology ,chemistry.chemical_compound ,Cell Line, Tumor ,Okadaic Acid ,Phosphoprotein Phosphatases ,Extracellular ,Humans ,Linoleic Acids, Conjugated ,Protein Phosphatase 2 ,Enzyme Inhibitors ,Extracellular Signal-Regulated MAP Kinases ,Mitogen-Activated Protein Kinase 1 ,Mitogen-Activated Protein Kinase 3 ,Nutrition and Dietetics ,Dose-Response Relationship, Drug ,integumentary system ,Cell growth ,Kinase ,food and beverages ,Okadaic acid ,Protein phosphatase 2 ,Molecular biology ,Culture Media ,Up-Regulation ,Cell biology ,chemistry ,MCF-7 ,Cell culture ,Carcinogens ,Female ,lipids (amino acids, peptides, and proteins) ,Cell Division - Abstract
Conjugated linoleic acid (CLA) has protective properties in breast cancer. Here, we studied the mechanisms underlying the effects of CLA on MCF-7 breast cancer cell proliferation, especially in correlation with the involvement of the extracellular signal-regulated kinase 1/2 (ERK1/2) pathway and protein phosphatase 2A (PP2A). CLA inhibits MCF-7 cell growth in a concentration- and time-dependent manner, without triggering apoptosis. In assessing expression levels of proteins that play obligatory roles in the ERK cascade, we evidenced that CLA down-regulated Raf-1 and decreased levels of phospho-ERK1/2, as well as c-myc expression. Increase in PP2A expression rates were additionally observed after CLA treatment of MCF-7 cells. The above effects, as well as CLA-induced inhibition of cell growth, were reversed by okadaic acid, a specific inhibitor of PP2A. Thus, PP2A likely participates in deactivation of ERK1/2, and its up-regulation may represent a novel mechanism for CLA-induced inhibition of cell proliferation.
- Published
- 2006
28. Arachidonic acid suppresses growth of human lung tumor A549 cells through down-regulation of ALDH3A1 expression
- Author
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Vasilis Vasiliou, Giuliana Muzio, Antonella Trombetta, Rosa Angela Canuto, Marina Maggiora, Germana Martinasso, and Natalie Lassen
- Subjects
medicine.medical_specialty ,Lung Neoplasms ,PPARγ ,Blotting, Western ,Lipid peroxidation ,Down-Regulation ,Aldehyde dehydrogenase ,Electrophoretic Mobility Shift Assay ,Free radicals ,Biology ,Biochemistry ,4-Hydroxynonenal ,chemistry.chemical_compound ,Downregulation and upregulation ,Cell Line, Tumor ,Physiology (medical) ,Internal medicine ,medicine ,Humans ,RNA, Messenger ,ALDH3A1 ,DNA Primers ,A549 human lung tumor cells ,A549 cell ,Base Sequence ,Reverse Transcriptase Polymerase Chain Reaction ,Human hepatoma cells ,Aldehyde Dehydrogenase ,PPAR gamma ,Endocrinology ,Arachidonic acid ,chemistry ,Cell culture ,biology.protein ,Growth inhibition ,Cell Division - Abstract
Expression of aldehyde dehydrogenase 3A1 (ALDH3A1) in certain normal and tumor cells is associated with protection against the growth inhibitory effect of reactive aldehydes generated during membrane lipid peroxidation. We found that human lung tumor (A549) cells, which express high levels of ALDH3A1 protein, were significantly less susceptible to the antiproliferative effects of 4-hydroxynonenal compared to human hepatoma HepG2 or SK-HEP-1 cells that lack ALDH3A1 expression. However, A549 cells became susceptible to lipid peroxidation products when they were treated with arachidonic acid. The growth suppression of A549 cells induced by arachidonic acid was associated with increased levels of lipid peroxidation and with reduced ALDH3A1 enzymatic activity, protein, and mRNA levels. Furthermore, arachidonic acid treatment of the A549 cells resulted in an increased expression of peroxisome proliferator-activated receptor gamma (PPARgamma), whereas NF-kappaB binding activity was inhibited. Blocking PPARgamma using a selective antagonist, GW9662, prevented the arachidonic acid-mediated reduction of ALDH3A1 expression as well as the growth inhibition of A549 cells, suggesting the central role of PPARgamma in these phenomena. The increase in PPARgamma and the reduction in ALDH3A1 were also prevented by exposing cells to vitamin E concomitant with arachidonic acid treatment. In conclusion, our data show that the arachidonic acid-induced suppression of A549 cell growth is associated with increased lipid peroxidation and decreased ALDH3A1 expression, which may be due to activation of PPARgamma.
- Published
- 2006
29. The Possible Role of Helicobacter pylori in Gastric Cancer and Its Management.
- Author
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Alfarouk, Khalid O., Bashir, Adil H. H., Aljarbou, Ahmed N., Ramadan, AbdelRahman M., Muddathir, Abdel Khalig, AlHoufie, Sari T. S., Hifny, Abdelhamid, Elhassan, Gamal O., Ibrahim, Muntaser E., Alqahtani, Saad S., AlSharari, Shakir D., Supuran, Claudiu T., Rauch, Cyril, Cardone, Rosa Angela, Reshkin, Stephan J., Fais, Stefano, and Harguindey, Salvador
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STOMACH cancer ,HELICOBACTER pylori ,ANAEROBIC bacteria ,CARCINOGENICITY ,BIOLOGY - Abstract
Helicobacter pylori (HP) is a facultative anaerobic bacterium. HP is a normal flora having immuno-modulating properties. This bacterium is an example of a microorganism inducing gastric cancer. Its carcinogenicity depends on bacteria-host related factors. The proper understanding of the biology of HP inducing gastric cancer offers the potential strategy in the managing of HP rather than eradicating it. In this article, we try to summarize the biology of HP-induced gastric cancer and discuss the current pharmacological approach to treat and prevent its carcinogenicity. [ABSTRACT FROM AUTHOR]
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- 2019
- Full Text
- View/download PDF
30. Na+/H+ Exchanger Regulatory Factor Isoform 1 Overexpression Modulates Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Expression and Activity in Human Airway 16HBE14o- Cells and Rescues ΔF508 CFTR Functional Expression in Cystic Fibrosis Cells*
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Stephan J. Reshkin, Salvatore Carrabino, Valeria Casavola, Stefania Maria Riccardi, Rosa Angela Cardone, Giovanni Busco, Edward J. Weinman, Maria Favia, Massimo Conese, Teresa Fanelli, Lorenzo Guerra, Dept Biosci Biotechnol & Bropharmaceut, Università degli Studi di Bari Aldo Moro, Centre de biophysique moléculaire (CBM), Université d'Orléans (UO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), Department of General and Environmental Physiology, Università degli studi di Bari Aldo Moro (UNIBA), Department of Biomedical Sciences, and Università degli Studi di Foggia - University of Foggia
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Cytoplasm ,Cystic Fibrosis ,Cell ,Cystic Fibrosis Transmembrane Conductance Regulator ,Gene Expression ,Biochemistry ,0302 clinical medicine ,Drug Stability ,Homeostasis ,Fluorescent Antibody Technique, Indirect ,ComputingMilieux_MISCELLANEOUS ,0303 health sciences ,Microscopy, Confocal ,biology ,Chemistry ,Transfection ,respiratory system ,Cystic fibrosis transmembrane conductance regulator ,Cell biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,congenital, hereditary, and neonatal diseases and abnormalities ,Sodium-Hydrogen Exchangers ,PDZ domain ,Bronchi ,Cell Line ,03 medical and health sciences ,Chlorides ,medicine ,Humans ,[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM] ,ΔF508 ,Protein kinase A ,Molecular Biology ,030304 developmental biology ,Binding Sites ,Cell Membrane ,Epithelial Cells ,Cell Biology ,Apical membrane ,Phosphoproteins ,Cyclic AMP-Dependent Protein Kinases ,Molecular biology ,digestive system diseases ,respiratory tract diseases ,Gene Expression Regulation ,Mutation ,biology.protein - Abstract
There is evidence that cystic fibrosis transmembrane conductance regulator (CFTR) interacting proteins play critical roles in the proper expression and function of CFTR. The Na(+)/H(+) exchanger regulatory factor isoform 1 (NHERF1) was the first identified CFTR-binding protein. Here we further clarify the role of NHERF1 in the regulation of CFTR activity in two human bronchial epithelial cell lines: the normal, 16HBE14o-, and the homozygous DeltaF508 CFTR, CFBE41o-. Confocal analysis in polarized cell monolayers demonstrated that NHERF1 distribution was associated with the apical membrane in 16HBE14o- cells while being primarily cytoplasmic in CFBE41o- cells. Transfection of 16HBE14o- monolayers with vectors encoding for wild-type (wt) NHERF1 increased both apical CFTR expression and apical protein kinase A (PKA)-dependent CFTR-mediated chloride efflux, whereas transfection with NHERF1 mutated in the binding groove of the PDZ domains or truncated for the ERM domain inhibited both the apical CFTR expression and the CFTR-dependent chloride efflux. These data led us to hypothesize an important role for NHERF1 in regulating CFTR localization and stability on the apical membrane of 16HBE14o- cell monolayers. Importantly, wt NHERF1 overexpression in confluent DeltaF508 CFBE41o- and DeltaF508 CFT1-C2 cell monolayers induced both a significant redistribution of CFTR from the cytoplasm to the apical membrane and a PKA-dependent activation of CFTR-dependent chloride secretion.
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- 2005
31. The role of disturbed pH dynamics and the Na+/H+ exchanger in metastasis
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Stephan J. Reshkin, Rosa Angela Cardone, and Valeria Casavola
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Sodium-Hydrogen Exchangers ,General Mathematics ,Cell ,Biology ,Mice ,Neoplasms ,medicine ,Extracellular ,Animals ,Humans ,Neoplasm Invasiveness ,Neoplasm Metastasis ,Cation Transport Proteins ,Sodium-Hydrogen Exchanger 1 ,Applied Mathematics ,Membrane Proteins ,Cell migration ,Hydrogen-Ion Concentration ,Cell biology ,Sodium–hydrogen antiporter ,medicine.anatomical_structure ,Cancer cell ,Invadopodia ,Signal transduction ,Cotransporter ,Signal Transduction - Abstract
Recent research has highlighted the fundamental role of the tumour's extracellular metabolic microenvironment in malignant invasion. This microenvironment is acidified primarily by the tumour-cell Na(+)/H(+) exchanger NHE1 and the H(+)/lactate cotransporter, which are activated in cancer cells. NHE1 also regulates formation of invadopodia - cell structures that mediate tumour cell migration and invasion. How do these alterations of the metabolic microenvironment and cell invasiveness contribute to tumour formation and progression?
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- 2005
32. Sesquiterpene lactones from Anthemis wiedemanniana
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Rosa Angela Raccuglia, Wanda Kisiel, Sergio Rosselli, Sezgin Çelik, Maurizio Bruno, İsmet Uysal, Antonella Maggio, CELIK S, ROSSELLI S, MAGGIO A, RACCUGLIA R A, UYSAL I, KISIEL W, and BRUNO M
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eudesmanolide ,biology ,Traditional medicine ,Asteraceae ,biology.organism_classification ,Sesquiterpene ,Biochemistry ,chemistry.chemical_compound ,chemistry ,sesquiterpene lactone ,Anthemis ,germacranolide ,Ecology, Evolution, Behavior and Systematics ,Anthemis wiedemanniana - Abstract
Asteraceae; Anthemis wiedemanniana; sesquiterpene lactones; germacranolides; eudesmanolide
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- 2005
33. Organization of phenylalanine ammonia lyase (PAL), acidic PR-5 and osmotin-like (OSM) defence-response gene families in the potato genome
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Marc Ghislain, Christiane Gebhardt, Carmen M. Herrera, and Rosa Angela Castillo Ruiz
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Chromosomes, Artificial, Bacterial ,DNA, Plant ,Sequence analysis ,Molecular Sequence Data ,Phenylalanine ammonia-lyase ,Biology ,Genome ,Gene mapping ,Genetics ,Gene family ,Amino Acid Sequence ,Molecular Biology ,Gene ,Phylogeny ,Phenylalanine Ammonia-Lyase ,Plant Proteins ,Solanum tuberosum ,Bacterial artificial chromosome ,Base Sequence ,Sequence Homology, Amino Acid ,fungi ,food and beverages ,General Medicine ,genomic DNA ,Multigene Family ,Genome, Plant - Abstract
Defence-response (DR) genes are candidates for the genetic functions underlying quantitative resistance to plant pathogens. The organization of three DR gene families encoding phenylalanine ammonia lyase (PAL), acidic PR-(pathogenesis-related) protein 5, and basic PR-5, or osmotin-like (OSM), proteins was studied in the potato genome. A bacterial artificial chromosome (BAC) library containing approximately 50,000 clones was constructed from high-molecular weight genomic DNA of the diploid potato clone PD59, a hybrid between Solanum tuberosum and S. phureja. BAC clones carrying one or more copies of the DR genes were identified and characterized by Southern hybridization, sequence analysis and genetic mapping. PAL, acidic PR-5 and OSM (basic PR-5) genes were all organized into gene families of varying complexity. The PAL gene family consisted of at least 16 members, several of which were physically linked. Four acidic PR-5 homologous were localized to a 45-kb segment on potato chromosome XII. One of these, PR-5/319, codes for the acidic thaumatin-like protein C found in intercellular fluids of potato. Nine OSM genes were organized at two loci: eight form a 90-kb cluster on chromosome VIII, and a single gene was found on chromosome XI. The topology of a phylogenetic tree based on PR-5 and OSM protein sequences from Solanaceae suggests a mode of evolution for these gene families. The results will form the basis for further studies on the potential role of these defence-related loci in quantitative resistance to pathogens.
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- 2005
34. Mechanisms involved in growth inhibition induced by clofibrate in hepatoma cells
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Giuliana Muzio, Sebastiano Colombatto, P. Reffo, Antonella Trombetta, Rosa Angela Canuto, Germana Martinasso, and Marina Maggiora
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MAPK/ERK pathway ,medicine.medical_specialty ,PPARgamma ,Receptors, Cytoplasmic and Nuclear ,Peroxisome proliferator-activated receptor ,Biology ,Toxicology ,Gene Expression Regulation, Enzymologic ,Oligodeoxyribonucleotides, Antisense ,Proto-Oncogene Proteins c-myc ,Liver Neoplasms, Experimental ,Internal medicine ,Okadaic Acid ,Phosphoprotein Phosphatases ,Tumor Cells, Cultured ,medicine ,Animals ,Protein Phosphatase 2 ,Clofibrate ,Enzyme Inhibitors ,Cell proliferation ,Mitogen-Activated Protein Kinase 1 ,chemistry.chemical_classification ,Hepatoma ,Mitogen-Activated Protein Kinase 3 ,L-Lactate Dehydrogenase ,Cell growth ,MAPK ,PP2A ,Protein phosphatase 2 ,Rats ,Cell biology ,Gene Expression Regulation, Neoplastic ,Endocrinology ,chemistry ,Apoptosis ,Cell culture ,Mitogen-Activated Protein Kinases ,Cell Division ,Intracellular ,Signal Transduction ,Transcription Factors ,medicine.drug - Abstract
Low concentrations of some peroxisome proliferators have been found to decrease apoptosis in rat liver cells, whereas higher but pharmacological concentrations have been found to inhibit cell proliferation or to induce apoptosis in human and rat hepatoma cells. The highly deviated JM2 rat hepatoma cell line was used to examine the mechanisms underlying the inhibitory effect on cell proliferation. Clofibrate chiefly inhibited cell proliferation in these cells. Parallel to the decrease in cell proliferation there was an increase of peroxisome proliferator activated receptor (PPAR) gamma and of protein phosphatase 2A, whose importance was confirmed, respectively, by using antisense oliginucleotides (AS-ODN) or okadaic acid. The increase of protein phosphatase 2A induced by PPARgamma caused a decrease of MAPK, an intracellular signaling transduction pathway, as shown by evaluation of Erk1,2 and c-myc. In light of these results, clofibrate, like conventional synthetic ligands of PPARgamma, may be regarded as a possible prototype anti-tumour drug.
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- 2003
35. Antibacterial Evaluation of Cnicin and Some Natural and Semisynthetic Analogues
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Felice Senatore, Francesco Napolitano, Rosa Angela Raccuglia, Maurizio Bruno, Sergio Rosselli, Antonella Maggio, M., Bruno, S., Rosselli, A., Maggio, R. A., Raccuglia, F., Napolitano, and Senatore, Felice
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Germacranolide ,Magnetic Resonance Spectroscopy ,Stereochemistry ,Pharmaceutical Science ,Centaurea ,Microbial Sensitivity Tests ,Biology ,Gram-Positive Bacteria ,Cnicin ,Analytical Chemistry ,chemistry.chemical_compound ,Gram-Negative Bacteria ,Drug Discovery ,Humans ,Organic chemistry ,Antibacterial agent ,Pharmacology ,Ester derivatives ,Organic Chemistry ,Salonitenolide ,Biological activity ,Anti-Bacterial Agents ,Complementary and alternative medicine ,chemistry ,Molecular Medicine ,Plant Preparations ,Antibacterial activity ,Sesquiterpenes ,Phytotherapy - Abstract
Some ester derivatives of salonitenolide have been prepared from cnicin, a germacranolide sesquiterpenoid. The compounds were tested for their antibacterial activity. The 8,15-diesters showed a good activity, comparable with that of cnicin. The 15-monoester compounds were not very active showing that esterification at the C-8 position is an important structural feature for antibacterial properties.
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- 2003
36. Apoptosis induced by clofibrate in Yoshida AH-130 hepatoma cells
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Rosa Angela Canuto, Marina Maggiora, Germana Martinasso, Paola Costelli, Antonella Trombetta, Giuliana Muzio, Francesco M. Baccino, Gabriella Bonelli, and Riccardo Autelli
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Clofibrate ,biology ,Cholesterol ,Farnesyltransferase ,cholesterol ,Cell Biology ,QD415-436 ,Reductase ,Biochemistry ,Cell biology ,chemistry.chemical_compound ,Cytosol ,mevalonate ,Endocrinology ,chemistry ,peroxisome proliferator activated receptors ,Apoptosis ,HMG-CoA reductase ,biology.protein ,medicine ,Protein farnesylation ,hydroxy-methyl-glutaryl coenzyme A reductase ,medicine.drug - Abstract
Clofibrate is a hypolipidemic drug belonging to the peroxisome proliferator (PP) family. PPs are well-recognized hepatocarcinogens, though only for rodents and not for humans. Their oncogenicity is usually ascribed to mitogenic or antiapoptotic action. However, we have reported that clofibrate can trigger fast and extensive apoptosis in rodent and human tumor cell lines. The present study examines the possible mechanisms involved in clofibrate-induced apoptosis in AH-130 hepatoma cells. The results show that the apoptogenic effect of clofibrate does not depend on induction of peroxisome proliferator activated receptors (PPARs), but on interference with HMG-CoA reductase (HMGR), a key enzyme that regulates cholesterol biosynthesis and production of isoprenoid units for protein farnesylation. The level and activity of HMGR mRNA are reduced in clofibrate-treated AH-130 cells and apoptosis can be partially prevented by addition of mevalonate. Moreover, cholesterol and cholesterol ester content decreases early in mitochondria, and cytocrome c is released in the cytosol. On the contrary, perturbations at the level of protein farnesylation are not important in determining the fast apoptogenic effect, since treatment of AH-130 cells with an inhibitor of farnesyltransferase induces apoptosis only after 4 h. In conclusion, inhibition of HMGR and decreased cholesterol content are crucial events in clofibrate-induced apoptosis in AH-130 hepatoma cells.
- Published
- 2003
37. Reciprocal Protein Kinase A Regulatory Interactions between Cystic Fibrosis Transmembrane Conductance Regulator and Na+/H+ Exchanger Isoform 3 in a Renal Polarized Epithelial Cell Model
- Author
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Stephan J. Reshkin, Lorenzo Guerra, Heini Murer, Valeria Casavola, Rosa Angela Cardone, Serge M. Gisler, Anna Bagorda, Teresa Fanelli, Francesca Di Sole, Corinna Hemle-Kolb, University of Zurich, and Casavola, V
- Subjects
Scaffold protein ,Cytoplasm ,1303 Biochemistry ,Time Factors ,Cystic Fibrosis Transmembrane Conductance Regulator ,Plasma protein binding ,Biochemistry ,10052 Institute of Physiology ,1307 Cell Biology ,0302 clinical medicine ,Ezrin ,Cyclic AMP ,Protein Isoforms ,Phosphorylation ,Glutathione Transferase ,0303 health sciences ,biology ,Kinase ,Hydrogen-Ion Concentration ,respiratory system ,Cystic fibrosis transmembrane conductance regulator ,3. Good health ,Cell biology ,Chlorine ,Protein Binding ,congenital, hereditary, and neonatal diseases and abnormalities ,Sodium-Hydrogen Exchangers ,Recombinant Fusion Proteins ,Blotting, Western ,Transfection ,Cell Line ,03 medical and health sciences ,1312 Molecular Biology ,Animals ,Biotinylation ,Protein kinase A ,Molecular Biology ,030304 developmental biology ,Ions ,Dose-Response Relationship, Drug ,urogenital system ,Epithelial Cells ,Cell Biology ,Oligonucleotides, Antisense ,Phosphoproteins ,Cyclic AMP-Dependent Protein Kinases ,Rats ,respiratory tract diseases ,Cytoskeletal Proteins ,Sodium–hydrogen antiporter ,Microscopy, Fluorescence ,Mutation ,biology.protein ,570 Life sciences ,Peptides ,030217 neurology & neurosurgery - Abstract
Although Cystic fibrosis transmembrane conductance regulator (CFTR) has been shown to regulate the activity of NHE3, the potential reciprocal interaction of NHE3 to modulate the protein kinase A (PKA)-dependent regulation of CFTR in epithelial cells is still unknown. In the present work, we describe experiments to define the interactions between CFTR and NHE3 with the regulatory, scaffolding protein, NHERF that organize their PKA-dependent regulation in a renal epithelial cell line that expresses endogenous CFTR. The expression of rat NHE3 significantly decreased PKA-dependent activation of CFTR without altering CFTR expression, and this decrease was prevented by mutation of either of the two rat NHE3 PKA target serines to alanine (S552A or S605A). Inhibition of CFTR expression by antisense treatment resulted in an acute decrease in PKA-dependent regulation of NHE3 activity. CFTR, NHE3, and ezrin were recognized by NHERF-2 but not NHERF-1 in glutathione S-transferase pull-down experiments. Ezrin may function as a protein kinase A anchoring protein (AKAP) in this signaling complex, because blocking the binding of PKA to an AKAP by incubation with the S-Ht31 peptide inhibited the PKA-dependent regulation of CFTR in the absence of NHE3. In the A6-NHE3 cells S-Ht31 blocked the PKA regulation of NHE3 whereas it now failed to affect the regulation of CFTR. We conclude that CFTR and NHE3 reciprocally interact via a shared regulatory complex comprised of NHERF-2, ezrin, and PKA.
- Published
- 2002
38. Key role of the expression of bone morphogenetic proteins in increasing the osteogenic activity of osteoblast-like cells exposed to shock waves and seeded on bioactive glass-ceramic scaffolds for bone tissue engineering
- Author
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Germana Martinasso, Roberto Frairia, Rosa Angela Canuto, Giuliana Muzio, Chiara Vitale-Brovarone, and Francesco Baino
- Subjects
Ceramics ,Materials science ,Bone Regeneration ,Osteocalcin ,Biomedical Engineering ,Bone morphogenetic protein ,Scaffold ,Bone morphogenetic proteins ,Shock waves ,Gremlin ,bone tissue engineering ,Bone and Bones ,law.invention ,Biomaterials ,law ,Osteogenesis ,Materials Testing ,medicine ,Humans ,RNA, Messenger ,Cell Proliferation ,Osteoblasts ,biology ,Tissue Engineering ,Tissue Scaffolds ,Osteoblast ,X-Ray Microtomography ,Alkaline Phosphatase ,Cell biology ,medicine.anatomical_structure ,Gene Expression Regulation ,Cell culture ,Bioactive glass ,Bone Morphogenetic Proteins ,biology.protein ,Alkaline phosphatase ,Gremlin (protein) ,Type I collagen ,Biomedical engineering - Abstract
In this work, the role of shock wave-induced increase of bone morphogenetic proteins in modulating the osteogenic properties of osteoblast-like cells seeded on a bioactive scaffold was investigated using gremlin as a bone morphogenetic protein antagonist. Bone-like glass-ceramic scaffolds, based on a silicate experimental bioactive glass developed at the Politecnico di Torino, were produced by the sponge replication method and used as porous substrates for cell culture. Human MG-63 cells, exposed to shock waves and seeded on the scaffolds, were treated with gremlin every two days and analysed after 20 days for the expression of osteoblast differentiation markers. Shock waves have been shown to induce osteogenic activity mediated by increased expression of alkaline phosphatase, osteocalcin, type I collagen, BMP-4 and BMP-7. Cells exposed to shock waves plus gremlin showed increased growth in comparison with cells treated with shock waves alone and, conversely, mRNA contents of alkaline phosphatase and osteocalcin were significantly lower. Therefore, the shock wave-mediated increased expression of bone morphogenetic protein in MG-63 cells seeded on the scaffolds is essential in improving osteogenic activity; blocking bone morphogenetic protein via gremlin completely prevents the increase of alkaline phosphatase and osteocalcin. The results confirmed that the combination of glass-ceramic scaffolds and shock waves exposure could be used to significantly improve osteogenesis opening new perspectives for bone regenerative medicine.
- Published
- 2014
39. Inhibition of cytosolic class 3 aldehyde dehydrogenase by antisense oligonucleotides in rat hepatoma cells
- Author
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Antonella Trombetta, Rosa Angela Canuto, Giuliana Muzio, and Marina Maggiora
- Subjects
Aldehyde dehydrogenase ,Antisense oligonucleotides ,Cell proliferation ,Hepatoma cells ,Lipid peroxidation ,Programmed cell death ,Cell division ,Gene Expression ,Endogeny ,Biology ,Toxicology ,Isozyme ,chemistry.chemical_compound ,Cytosol ,Liver Neoplasms, Experimental ,Tumor Cells, Cultured ,Animals ,RNA, Messenger ,RNA, Neoplasm ,Enzyme Inhibitors ,Cell growth ,General Medicine ,Aldehyde Dehydrogenase ,Oligonucleotides, Antisense ,Molecular biology ,Rats ,Biochemistry ,chemistry ,biology.protein ,Cell Division - Abstract
Aldehyde dehydrogenases (ALDHs) are a superfamily of several isoenzymes widely expressed in bacteria, yeast, plant and animals. Three major classes of ALDHs have been traditionally identified, classes 1, 2 and 3. Both exogenous and endogenous aldehydes, including aldehydes derived from lipid peroxidation, are oxidized by the ALDH superfamily. Several changes in ALDH isoenzyme expression take place in hepatoma cells, in particular cytosolic class 3 ALDH (ALDH3), not expressed in normal hepatocytes, appears and increases with the degree of deviation. It has been demonstrated that cytosolic ALDH3 is important in determining the resistance of tumor cells to antitumor drugs, such as cyclophosphamide. Moreover, hepatoma-associated ALDH3 seems to be important in metabolizing aldehydes derived from lipid peroxidation, and in particular the cytostatic aldehyde 4-hydroxynonenal (4-HNE). We demonstrated previously that restoring endogenous lipid peroxidation in hepatoma cells by enriching them with arachidonic acid causes a decrease of mRNA, protein and enzyme activity of ALDH3 and that this decrease reduces cell growth and/or causes cell death, depending on basal class 3 ALDH activity. To confirm the correlation between inhibition of class 3 ALDH and reduction of cell proliferation, we exposed hepatoma cells to antisense oligonucleotides (ODNs) against ALDH3. In JM2 hepatoma cell line, with high ALDH3 activity, the exposure to antisense ODNs significantly decreases mRNA and enzyme activity (90%). At the same time, cell growth was reduced by about 70%. The results confirm that in hepatoma cells ALDH3 expression is closely related with cell growth, and that its inhibition is important in reducing the proliferation of hepatoma cells overexpressing ALDH3.
- Published
- 2001
40. Dose-dependent inhibition of cell proliferation induced by lipid peroxidation products in rat hepatoma cells after enrichment with arachidonic acid
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Mario U. Dianzani, Mario Terreno, Rosa Angela Canuto, Riccardo Autelli, Raffaella A. Salvo, Marina Maggiora, Antonella Trombetta, and Giuliana Muzio
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Clinical chemistry ,Aldehyde dehydrogenase ,Biochemistry ,Lipid peroxidation ,chemistry.chemical_compound ,Liver Neoplasms, Experimental ,Tumor Cells, Cultured ,Animals ,chemistry.chemical_classification ,Arachidonic Acid ,Dose-Response Relationship, Drug ,L-Lactate Dehydrogenase ,biology ,Cell growth ,Organic Chemistry ,Cell Biology ,Enzyme assay ,Culture Media ,Rats ,chemistry ,biology.protein ,lipids (amino acids, peptides, and proteins) ,Arachidonic acid ,Lipid Peroxidation ,Signal transduction ,Cell Division ,Polyunsaturated fatty acid - Abstract
Polyunsaturated fatty acids (PUFA) are important constituents of membrane phospholipids, whose levels are decreased in some tumor cells. This deficiency may cause alterations in signal transduction and an interruption of normal cellular events. The enrichment of tumor cells with PUFA may stimulate or inhibit tumor growth, probably depending on the type of PUFA and the cellular concentration of aldehydes derived from restored lipid peroxidation. We examined the effect of several doses of prooxidant on the growth of hepatoma cells with different aldehyde dehydrogenase activities, enriched with arachidonic acid. Two doses of prooxidant were sufficient to reduce growth of hepatoma cells with low aldehyde dehydrogenase activity, whereas three doses were necessary for those with high enzyme activity. In both cases, lipid peroxidation products blocked the cells in the S phase.
- Published
- 1999
41. Vitrification preserves chromatin integrity, bioenergy potential and oxidative parameters in mouse embryos
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Nicola Antonio Martino, Maria Elena Dell'Aquila, Bence Somoskoi, Giovanni Michele Lacalandra, Sándor Cseh, and Rosa Angela Cardone
- Subjects
Male ,Ethylene Glycol ,animal structures ,Cryoprotectant ,Biology ,medicine.disease_cause ,Morula ,Cryopreservation ,Andrology ,Mice ,Cryoprotective Agents ,Endocrinology ,Pregnancy ,Botany ,medicine ,Animals ,Vitrification ,Blastocyst ,Research ,Embryogenesis ,Reproducibility of Results ,Obstetrics and Gynecology ,Embryo ,Blastomere ,Embryo, Mammalian ,Propylene Glycol ,Chromatin ,Mitochondria ,medicine.anatomical_structure ,Reproductive Medicine ,embryonic structures ,Female ,Energy Metabolism ,Reactive Oxygen Species ,Oxidation-Reduction ,Oxidative stress ,Developmental Biology - Abstract
Background The aim of this study was to evaluate the effects of vitrification on morpho-functional parameters (blastomere/chromatin integrity and bioenergy/oxidative potential) of mouse preimplantation embryos. Methods In vivo produced mouse (4/16-cell, morulae and blastocyst-stage) embryos were randomly divided into vitrification and control groups. For vitrification, embryos were exposed to a 2-step loading of ethylene glycol and propylene glycol, before being placed in a small nylon loop and submerged into liquid nitrogen. After warming, the cryoprotectants were diluted by a 3-step procedure. Embryo morphology, chromatin integrity and energy/oxidative status were compared between groups. Results Vitrification induced low grade blastomere cytofragmentation (P
- Published
- 2013
42. ß1 integrin binding phosphorylates ezrin at T567 to activate a lipid raft signalsome driving invadopodia activity and invasion
- Author
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Francesca Di Sole, MariaLuisa L. Carcangiu, Loredana Moro, Ester Antelmi, Rosa Rubino, Nicola Antonio Martino, Stephan J. Reshkin, Valeria Casavola, Rosa Angela Cardone, and Maria Raffaella Greco
- Subjects
genetic structures ,Receptor, ErbB-2 ,Science ,Integrin ,Fluorescent Antibody Technique ,Breast Neoplasms ,macromolecular substances ,Biology ,environment and public health ,Ezrin ,Membrane Microdomains ,Image Processing, Computer-Assisted ,Humans ,Immunoprecipitation ,Neoplasm Invasiveness ,Pseudopodia ,Phosphorylation ,Cytoskeleton ,Lipid raft ,Integrin binding ,DNA Primers ,Analysis of Variance ,Multidisciplinary ,Integrin beta1 ,Actin cytoskeleton ,Immunohistochemistry ,Cell biology ,Extracellular Matrix ,Cytoskeletal Proteins ,Gene Expression Regulation ,Italy ,Invadopodia ,biology.protein ,Medicine ,Female ,Signal Transduction ,Research Article - Abstract
Extracellular matrix (ECM) degradation is a critical process in tumor cell invasion and requires matrix degrading protrusions called invadopodia. The Na(+)/H(+) exchanger (NHE1) has recently been shown to be fundamental in the regulation of invadopodia actin cytoskeleton dynamics and activity. However, the structural link between the invadopodia cytoskeleton and NHE1 is still unknown. A candidate could be ezrin, a linker between the NHE1 and the actin cytoskeleton known to play a pivotal role in invasion and metastasis. However, the mechanistic basis for its role remains unknown. Here, we demonstrate that ezrin phosphorylated at T567 is highly overexpressed in the membrane of human breast tumors and positively associated with invasive growth and HER2 overexpression. Further, in the metastatic cell line, MDA-MB-231, p-ezrin was almost exclusively expressed in invadopodia lipid rafts where it co-localized in a functional complex with NHE1, EGFR, ß1-integrin and phosphorylated-NHERF1. Manipulation by mutation of ezrins T567 phosphorylation state and/or PIP2 binding capacity or of NHE1s binding to ezrin or PIP2 demonstrated that p-ezrin expression and binding to PIP2 are required for invadopodia-mediated ECM degradation and invasion and identified NHE1 as the membrane protein that p-ezrin regulates to induce invadopodia formation and activity.
- Published
- 2013
43. CFTR regulation in human airway epithelial cells requires integrity of the actin cytoskeleton and compartmentalized cAMP and PKA activity
- Author
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Rosa Angela Cardone, Domenico Marzulli, Maria Favia, Stefania Monterisi, Lorenzo Guerra, Manuela Zaccolo, Stephan J. Reshkin, and Valeria Casavola
- Subjects
Cytoplasm ,Sodium-Hydrogen Exchangers ,Cytoskeleton organization ,Cystic Fibrosis Transmembrane Conductance Regulator ,Respiratory Mucosa ,Biology ,Cystic fibrosis ,Cell Line ,NHERF1 ,cAMP ,Cyclic AMP ,Humans ,PKA ,RNA, Small Interfering ,Cytoskeleton ,Research Articles ,Epithelial Cells ,Cell Biology ,Compartmentalization (psychology) ,Apical membrane ,Actin cytoskeleton ,Phosphoproteins ,Cyclic AMP-Dependent Protein Kinases ,Cystic fibrosis transmembrane conductance regulator ,Cell biology ,Actin Cytoskeleton ,Cytoskeletal Proteins ,airways cells ,biology.protein ,RNA Interference ,Signal transduction ,Intracellular ,Signal Transduction - Abstract
The cystic fibrosis transmembrane conductance regulator (CFTR) mutation ΔF508CFTR still causes regulatory defects when rescued to the apical membrane, suggesting that the intracellular milieu might affect its ability to respond to cAMP regulation. We recently reported that overexpression of the Na+/H+ exchanger regulatory factor NHERF1 in the cystic fibrosis (CF) airway cell line CFBE41o-rescues the functional expression of ΔF508CFTR by promoting F-actin organization and formation of the NHERF1–ezrin–actin complex. Here, using real-time FRET reporters of both PKA activity and cAMP levels, we find that lack of an organized subcortical cytoskeleton in CFBE41o-cells causes both defective accumulation of cAMP in the subcortical compartment and excessive cytosolic accumulation of cAMP. This results in reduced subcortical levels and increased cytosolic levels of PKA activity. NHERF1 overexpression in CFBE41o-cells restores chloride secretion, subcortical cAMP compartmentalization and local PKA activity, indicating that regulation of ΔF508CFTR function requires not only stable expression of the mutant CFTR at the cell surface but also depends on both generation of local cAMP signals of adequate amplitude and activation of PKA in proximity of its target. Moreover, we found that the knockdown of wild-type CFTR in the non-CF 16HBE14o-cells results in both altered cytoskeletal organization and loss of cAMP compartmentalization, whereas stable overexpression of wt CFTR in CF cells restores cytoskeleton organization and re-establishes the compartmentalization of cAMP at the plasma membrane. This suggests that the presence of CFTR on the plasma membrane influences the cytoskeletal organizational state and, consequently, cAMP distribution. Our data show that a sufficiently high concentration of cAMP in the subcortical compartment is required to achieve PKA-mediated regulation of CFTR activity.
- Published
- 2012
44. Aldehyde dehydrogenases and cell proliferation
- Author
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Marina Maggiora, Elena Paiuzzi, Rosa Angela Canuto, Giuliana Muzio, and Manuela Oraldi
- Subjects
PPARs ,Peroxisome proliferator-activated receptor ,Aldehyde dehydrogenase ,Stem cells ,Biology ,Biochemistry ,4-Hydroxynonenal ,aldehyde dehydrogenase ,cancer ,PUFAs ,chemistry.chemical_compound ,Cancer stem cell ,Physiology (medical) ,Neoplasms ,Animals ,Humans ,Enzyme Inhibitors ,Antineoplastic Agents, Alkylating ,ALDH2 ,Cell Proliferation ,chemistry.chemical_classification ,Cell growth ,Transfection ,Peroxisome ,Enzyme Activation ,Oxidative Stress ,chemistry ,Drug Resistance, Neoplasm ,biology.protein ,Lipid Peroxidation - Abstract
Aldehyde dehydrogenases (ALDHs) oxidize aldehydes to the corresponding carboxylic acids using either NAD or NADP as a coenzyme. Aldehydes are highly reactive aliphatic or aromatic molecules that play an important role in numerous physiological, pathological, and pharmacological processes. ALDHs have been discovered in practically all organisms and there are multiple isoforms, with multiple subcellular localizations. More than 160 ALDH cDNAs or genes have been isolated and sequenced to date from various sources, including bacteria, yeast, fungi, plants, and animals. The eukaryote ALDH genes can be subdivided into several families; the human genome contains 19 known ALDH genes, as well as many pseudogenes. Noteworthy is the fact that elevated activity of various ALDHs, namely ALDH1A2, ALDH1A3, ALDH1A7, ALDH2*2, ALDH3A1, ALDH4A1, ALDH5A1, ALDH6, and ALDH9A1, has been observed in normal and cancer stem cells. Consequently, ALDHs not only may be considered markers of these cells, but also may well play a functional role in terms of self-protection, differentiation, and/or expansion of stem cell populations. The ALDH3 family includes enzymes able to oxidize medium-chain aliphatic and aromatic aldehydes, such as peroxidic and fatty aldehydes. Moreover, these enzymes also have noncatalytic functions, including antioxidant functions and some structural roles. The gene of the cytosolic form, ALDH3A1, is localized on chromosome 17 in human beings and on the 11th and 10th chromosome in the mouse and rat, respectively. ALDH3A1 belongs to the phase II group of drug-metabolizing enzymes and is highly expressed in the stomach, lung, keratinocytes, and cornea, but poorly, if at all, in normal liver. Cytosolic ALDH3 is induced by polycyclic aromatic hydrocarbons or chlorinated compounds, such as 2,3,7,8-tetrachlorodibenzo-p-dioxin, in rat liver cells and increases during carcinogenesis. It has been observed that this increased activity is directly correlated with the degree of deviation in hepatoma and lung cancer cell lines, as is the case in chemically induced hepatoma in rats. High ALDH3A1 expression and activity have been correlated with cell proliferation, resistance against aldehydes derived from lipid peroxidation, and resistance against drug toxicity, such as oxazaphosphorines. Indeed, cells with a high ALDH3A1 content are more resistant to the cytostatic and cytotoxic effects of lipidic aldehydes than are those with a low content. A reduction in cell proliferation can be observed when the enzyme is directly inhibited by the administration of synthetic specific inhibitors, antisense oligonucleotides, or siRNA or indirectly inhibited by the induction of peroxisome proliferator-activated receptor γ (PPARγ) with polyunsaturated fatty acids or PPARγ transfection. Conversely, cell proliferation is stimulated by the activation of ALDH3A1, whether by inhibiting PPARγ with a specific antagonist, antisense oligonucleotides, siRNA, or a medical device (i.e., composite polypropylene prosthesis for hernia repair) used to induce cell proliferation. To date, the mechanisms underlying the effects of ALDHs on cell proliferation are not yet fully clear. A likely hypothesis is that the regulatory effect is mediated by the catabolism of some endogenous substrates deriving from normal cell metabolism, such as 4-hydroxynonenal, which have the capacity to either stimulate or inhibit the expression of genes involved in regulating proliferation.
- Published
- 2012
45. E6 and E7 from human papillomavirus type 16 cooperate to target the PDZ protein Na/H exchange regulatory factor 1
- Author
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Bakary S. Sylla, Miranda Thomas, Lawrence Banks, Massimo Tommasino, Mariafrancesca Scalise, Rosa Rubino, Naveed Shahzad, Cesare Indiveri, Tarik Gheit, Rosa Angela Cardone, Stephan J. Reshkin, and Rosita Accardi
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Sodium-Hydrogen Exchangers ,Guanylate kinase ,Papillomavirus E7 Proteins ,Immunology ,PDZ domain ,Immunoblotting ,PDZ Domains ,Plasma protein binding ,Biology ,Microbiology ,Mice ,Phosphatidylinositol 3-Kinases ,Virology ,Animals ,Humans ,Gene Silencing ,Phosphorylation ,RNA, Small Interfering ,PI3K/AKT/mTOR pathway ,Human papillomavirus 16 ,Kinase ,Akt/PKB signaling pathway ,Reverse Transcriptase Polymerase Chain Reaction ,virus diseases ,Oncogene Proteins, Viral ,Phosphoproteins ,Molecular biology ,female genital diseases and pregnancy complications ,Cell biology ,Virus-Cell Interactions ,Repressor Proteins ,HEK293 Cells ,Insect Science ,NIH 3T3 Cells ,Signal transduction ,Protein Binding ,Signal Transduction - Abstract
Previous studies have shown that the PDZ-binding motif of the E6 oncoprotein from the mucosal high-risk (HR) human papillomavirus (HPV) types plays a key role in HPV-mediated cellular transformation in in vitro and in vivo experimental models. HR HPV E6 oncoproteins have the ability to efficiently degrade members of the PDZ motif-containing membrane-associated guanylate kinase (MAGUK) family; however, it is possible that other PDZ proteins are also targeted by E6. Here, we describe a novel interaction of HPV type 16 (HPV16) E6 with a PDZ protein, Na + /H + exchange regulatory factor 1 (NHERF-1), which is involved in a number of cellular processes, including signaling and transformation. HPV16 E6 associates with and promotes the degradation of NHERF-1, and this property is dependent on the C-terminal PDZ-binding motif of E6. Interestingly, HPV16 E7, via the activation of the cyclin-dependent kinase complexes, promoted the accumulation of a phosphorylated form of NHERF-1, which is preferentially targeted by E6. Thus, both oncoproteins appear to cooperate in targeting NHERF-1. Notably, HPV18 E6 is not able to induce NHERF-1 degradation, indicating that this property is not shared with E6 from all HR HPV types. Downregulation of NHERF-1 protein levels was also observed in HPV16-positive cervical cancer-derived cell lines, such as SiHa and CaSki, as well as HPV16-positive cervical intraepithelial neoplasia (CIN). Finally, our data show that HPV16-mediated NHERF-1 degradation correlates with the activation of the phosphatidylinositol-3′-OH kinase (PI3K)/AKT signaling pathway, which is known to play a key role in carcinogenesis.
- Published
- 2011
46. Cytotoxic geranylflavonoids from Bonannia graeca
- Author
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Sergio Rosselli, Kenneth F. Bastow, Rosa Angela Raccuglia, Muhammad Safder, Kuo Hsiung Lee, Chin Yu Lai, Antonella Maggio, Maurizio Bruno, Rosselli, S, Bruno, M, Maggio, A, Raccuglia, R A, Safder, M, Lai, C Y, Bastow, K F, and Lee, K H
- Subjects
Synthetic derivatives ,Stereochemistry ,Chemical structure ,Plant Science ,Horticulture ,Biochemistry ,Article ,Structure-Activity Relationship ,Bonannia graeca ,Cell Line, Tumor ,Humans ,Cytotoxic T cell ,Structure–activity relationship ,Settore BIO/15 - Biologia Farmaceutica ,Geranylflavonoids ,Molecular Biology ,Flavonoids ,Apiaceae ,Molecular Structure ,biology ,Cytotoxic activity ,fungi ,Euphorbiaceae ,food and beverages ,General Medicine ,Settore CHIM/06 - Chimica Organica ,Plant Components, Aerial ,biology.organism_classification ,Antineoplastic Agents, Phytogenic ,Human tumor ,Two-dimensional nuclear magnetic resonance spectroscopy - Abstract
The analysis of the aerial parts of Bonannia graeca led to the isolation and characterization of two new polar geranylated flavonoids (6 and 7). The structure elucidation was performed by extensive spectroscopic methods (1D and 2D NMR) and comparison with literature data. All natural flavonoids isolated from B. graeca (1–7) and some synthetic derivatives (8–11) were tested for cytotoxic activity against four human tumor cell lines. Preliminary structure-activity relationship correlations are discussed.
- Published
- 2011
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47. Tissue protein turnover during liver carcinogenesis
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Francesco M. Baccino, Giuliana Muzio, Luciana Tessitore, Riccardo Autelli, and Rosa Angela Canuto
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Male ,Cancer Research ,medicine.medical_specialty ,Cell division ,Cell ,Protein degradation ,Biology ,medicine.disease_cause ,Liver Neoplasms, Experimental ,In vivo ,Internal medicine ,medicine ,Animals ,Diethylnitrosamine ,Cell growth ,Protein turnover ,General Medicine ,medicine.disease ,Rats, Inbred F344 ,Neoplasm Proteins ,Rats ,medicine.anatomical_structure ,Endocrinology ,Liver ,Cancer research ,Liver cancer ,Carcinogenesis ,Precancerous Conditions ,Cell Division - Abstract
Overall rates of tissue protein degradation in vivo during chemical hepatocarcinogenesis were estimated by a double-isotope method as well as from the accumulation of peptide intermediates in protein degradation induced by bestatin. Several parameters estimating rates of cell proliferation and cell loss have been measured in parallel. The two procedures adopted consistently indicated that protein turnover was significantly slowed down through the whole observation period (12 months after the initiating administration of DENA) in both 'preneoplastic' nodules and hepatomas as compared with control livers or perinodular tissue. Such a difference may confer a selective growth advantage to 'preneoplastic' and tumoral cells. Since protein degradation rates did not appreciably differ between nodules and hepatomas, either such advantage originated from some early step in the carcinogenetic process or it merely reflected the proliferative events in the two cell populations. Yet neither liver nodules nor hepatomas were characterized by very high rates of cell proliferation, however much increased with respect to control liver.
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- 1993
48. NHERF1/EBP50 in Breast Cancer: Clinical Perspectives
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Anita Mangia, Antonia Bellizzi, Andrea Malfettone, and Rosa Angela Cardone
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biology ,Tumor hypoxia ,business.industry ,PDZ domain ,Review Article ,medicine.disease ,Bioinformatics ,HER2/neu ,Breast cancer ,Oncology ,Cell surface receptor ,Cancer research ,medicine ,biology.protein ,Carcinoma ,Surgery ,Receptor ,business ,Postsynaptic density - Abstract
Na(+)/H(+) exchanger regulatory factor 1 (NHERF1) is a postsynaptic density 95/disc-large/zona occludens (PDZ) domain-containing protein that recruits membrane receptors/transporters and cytoplasmic signaling proteins into functional complexes. NHERF1 expression has been demonstrated to be altered in breast cancer, but its role in mammary cancerogenesis and progression remains still undefined. In this paper, we review what is known on the pathological role and the potential clinical application of NHERF1 protein in breast cancer. Recent evidence shows that an increased cytoplasmic expression of NHERF1 suggests a key role of its localization/compartmentalization in defining cancerogenesis, progression, and invasion. NHERF1 overexpression is associated with increasing tumor cytohistological grade, aggressive clinical behavior, unfavorable prognosis, and increased tumor hypoxia. Moreover, NHERF1 co-localizes with the oncogenic receptor HER2/neu in HER2/neu-overexpressing carcinoma and in distant metastases. These data make NHERF1 also a potential candidate of clinical relevance for anti-HER2/neu therapy.
- Published
- 2010
49. Involvement of PPARα and PPARγ in apoptosis and proliferation of human hepatocarcinoma HepG2 cells
- Author
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Marina Maggiora, Rosa Angela Canuto, Giuliana Muzio, and Manuela Oraldi
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medicine.medical_specialty ,Carcinoma, Hepatocellular ,Time Factors ,PPARs ,medicine.drug_class ,Hepatoma cells ,Clinical Biochemistry ,Peroxisome proliferator-activated receptor ,Clofibrate ,PGJ2 ,Apoptosis ,Fibrate ,Biology ,Ligands ,Biochemistry ,Proto-Oncogene Proteins c-myc ,Internal medicine ,medicine ,Humans ,PPAR alpha ,Protein Phosphatase 2 ,Receptor ,Cell Proliferation ,chemistry.chemical_classification ,Cell growth ,Prostaglandin D2 ,Cell Cycle ,Liver Neoplasms ,Osmolar Concentration ,Cell Biology ,General Medicine ,Hep G2 Cells ,Cell cycle ,PPAR gamma ,Endocrinology ,chemistry ,Cancer research ,bcl-Associated Death Protein ,Signal transduction ,medicine.drug ,Signal Transduction - Abstract
Peroxisome proliferator-activated receptors (PPARs) mediate the effects of various ligands, known as peroxisome proliferators, a heterogeneous class of compounds including industrial chemicals, pharmaceuticals, and biomolecules such as fatty acids and eicosanoids. Among peroxisome proliferators, fibrate derivatives are considered specific ligands for PPARα, whereas eicosanoids, such as PGJ2, for PPARγ. The study aimed to clarify the relation between PPARs and apoptosis or proliferation on the same type of cells, using clofibrate as specific ligand of PPARα and PGJ2 as specific ligand of PPARγ. The cells used were human hepatocarcinoma HepG2 cells. The results showed that PPARα protein content increased in HepG2 cells treated with clofibrate, causing apoptosis in a time- and concentration-dependent way, as evidenced by the citofluorimetric assay and determination of BAD, myc and protein phosphatase 2A protein content. It also emerged that PPARγ increased in the same cells when treated with a specific ligand of this PPAR; in this case the increase of PPARγ did not cause an increase of apoptosis, but a time- and concentration-dependent inhibition of cell proliferation, evidenced by decreased cell numbers and increased number of cells in the G0/G1 phase of the cycle. It may be concluded that PPARα is chiefly related to apoptosis and PPARγ to cell proliferation.
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- 2010
50. NHE1 promotes invadopodial ECM proteolysis through acidification of the peri-invadopodial space
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Maria Elena Dell'Aquila, Angelo Paradiso, Maria Raffaella Greco, Ester Antelmi, Giovanni Busco, Maria Teresa Mancini, Stephan J. Reshkin, Valeria Casavola, Rosa Angela Cardone, Antonia Bellizzi, Matilde Colella, Centre de biophysique moléculaire (CBM), Université d'Orléans (UO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), and University of Pisa - Università di Pisa
- Subjects
Proteases ,Sodium-Hydrogen Exchangers ,Podosome ,Proteolysis ,Guinea Pigs ,[SDV.CAN]Life Sciences [q-bio]/Cancer ,Biology ,Biochemistry ,Extracellular matrix ,03 medical and health sciences ,0302 clinical medicine ,Osteoclast ,RNA interference ,Genetics ,medicine ,Animals ,Humans ,Secretion ,[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM] ,Molecular Biology ,ComputingMilieux_MISCELLANEOUS ,030304 developmental biology ,0303 health sciences ,medicine.diagnostic_test ,Hydrolysis ,Anatomy ,Cell biology ,Extracellular Matrix ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Invadopodia ,Biotechnology - Abstract
Extracellular matrix (ECM) degradation is a critical process in tumor cell invasion and requires membrane and released proteases focalized at membrane structures called invadopodia. While extracellular acidification is important in driving tumor invasion, the structure/function mechanisms underlying this regulation are still unknown. Invadopodia are similar in structure and function to osteoclast podosomes responsible for bone degradation, and extracellular acidification is central to podosome action, suggesting that it could also be for invadopodial function. Here, utilizing a novel system for in situ zymography in native matrices, we show that the Na(+)/H(+) exchanger (NHE1) and NHE1-generated extracellular acidification are localized at and necessary for invadopodial-dependent ECM degradation, thereby promoting tumor invasion. Stimulation with EGF increased both NHE1-dependent proton secretion and ECM degradation. Manipulation of the NHE1 expression by RNA interference or activity via either transport-deficient mutation or the specific inhibitor cariporide confirmed that NHE1 expression and activity are required for invadopodia-mediated ECM degradation. Taken together, our data show a concordance among NHE1 localization, the generation of a well-defined acidic extracellular pH in the nanospace surrounding invadopodia, and matrix-degrading activity at invadopodia of human malignant breast carcinoma cells, providing a structural basis for the role of NHE1 in invasion and identifying NHE1 as a strategic target for therapeutic intervention.
- Published
- 2010
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