title: Effects of nitric oxide synthase-3 overexpression on post-translational modifications and cell survival in HepG2 cells.
abstract: Hepatocarcinoma is the fifth most common neoplasm and the third cause of cancer-related death. The development of genetic- and/or molecular-based therapies is urgently required. The administration of high doses of nitric oxide (NO) promotes cell death in hepatocytes. NO contributes to cell signaling by inducing oxidative/nitrosative-dependent post-translational modifications. The aim of the present study was to investigate protein modifications and its relation with alteration of cell proliferation and death in hepatoma cells. Increased intracellular NO production was achieved by stable nitric oxide synthase-3 (NOS-3) overexpression in HepG2 cells. We assessed the pattern of nitration, nitrosylation and carbonylation of proteins by proteomic analysis. The results showed that NOS-3 cell overexpression increased oxidative stress, which affected proteins mainly involved in cell protein folding. Carbonylation also altered metabolism, as well as immune and antioxidant responses. The interaction of nitrosative and oxidative stress generated tyrosine nitration, which affected the tumor marker Serpin B3, ATP synthesis and cytoskeleton. All these effects were associated with a decrease in chaperone activity, a reduction in cell proliferation and an increased cell death. Our study showed that alteration of nitration, nitrosylation and carbonylation pattern of proteins by NO-dependent oxidative/nitrosative stress was related to a reduction of cell survival in a hepatoma cell line.
authors.affiliation: Liver Research Unit, IMIBIC (Instituto Maimónides para la Investigación Biomédica de Córdoba), Reina Sofia University Hospital, Córdoba, Spain. b92agmep@uco.es
authors.forename: P
authors.initials: P
authors.lastname: Aguilar-Melero
authors.forename: G
authors.initials: G
authors.lastname: Ferrín
authors.forename: J
authors.initials: J
authors.lastname: Muntané
mesh_terms: D000255:Adenosine Triphosphate / Q000096:biosynthesis / Q000235:genetics; D000951:Antigens, Neoplasm / Q000096:biosynthesis / Q000235:genetics; D006528:Carcinoma, Hepatocellular / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D016923:Cell Death / Q000235:genetics; D049109:Cell Proliferation; D002470:Cell Survival; D003599:Cytoskeleton / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D009363:Neoplasm Proteins / Q000096:biosynthesis* / Q000235:genetics; D009569:Nitric Oxide / Q000096:biosynthesis / Q000235:genetics; D052250:Nitric Oxide Synthase Type III / Q000096:biosynthesis* / Q000235:genetics; D018384:Oxidative Stress / Q000235:genetics; D017510:Protein Folding; D011499:Protein Processing, Post-Translational; D015843:Serpins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D000951:Antigens, Neoplasm; D009363:Neoplasm Proteins; D015843:Serpins; C051072:squamous cell carcinoma-related antigen; D009569:Nitric Oxide; D000255:Adenosine Triphosphate; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III