title: Redox regulation of p53 during hypoxia.
abstract: The transcription factor p53 can induce growth arrest or death in cells. Tumor cells that develop mutations in p53 demonstrate a diminished apoptotic potential, which may contribute to growth and tumor metastasis. Cellular levels of p53 are stabilized during hypoxia. The present study tested the hypothesis that reactive oxygen species (ROS) released from mitochondria regulate the cytosolic redox state and are required for the stabilization of p53 protein levels in response to hypoxia. Our results indicate that hypoxia (1.5% O2) increases mitochondrial ROS generation and increases p53 protein levels in human breast carcinoma MCF-7 cells and in normal human diploid fibroblast IMR-90 cells. MCF-7 cells depleted of their mitochondrial DNA (rho(o) cells) failed to stabilize p53 protein levels during hypoxia. The antioxidant N-acetylcysteine and the Cu/Zn superoxide dismutase inhibitor diethyldithiocarbamic acid abolished the hypoxia-induced increases in ROS and p53 levels. Rotenone, an inhibitor of mitochondrial complex I, and 4,4'-diisothiocyanato-stilbene-2,2'-disulfonate, a mitochondrial anion channel inhibitor, also abolished the increase in ROS signal and p53 levels during hypoxia. The p53-dependent gene p21WAF1/CIP1 was also induced by hypoxia in both MCF-7 and IMR-90 cells without affecting the growth rate of either cell line. In contrast, both cell lines exhibited increases in p21WAF1/CIP1 expression and growth arrest after gamma irradiation. Primary chick cardiac myocytes and murine embryonic fibroblasts also showed an increase in p53 protein levels in response to hypoxia without cell death or growth arrest. These results indicate that mitochondria regulate p53 protein levels during hypoxia through a redox-dependent mechanism involving ROS. Despite p53-induction, hypoxia alone does not cause either growth arrest or cell death.
authors.affiliation: Department of Medicine, Gwen Knapp Center, Committee on Immunology and the Howard Hughes Medical Institute, The University of Chicago, Illinois, USA.
authors.forename: N S
authors.initials: NS
authors.lastname: Chandel
authors.forename: M G
authors.initials: MG
authors.lastname: Vander Heiden
authors.forename: C B
authors.initials: CB
authors.lastname: Thompson
authors.forename: P T
authors.initials: PT
authors.lastname: Schumacker
mesh_terms: D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid / Q000494:pharmacology; D000111:Acetylcysteine / Q000494:pharmacology; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D001943:Breast Neoplasms / Q000378:metabolism; D002277:Carcinoma / Q000378:metabolism; D002453:Cell Cycle / Q000528:radiation effects; D016923:Cell Death / Q000187:drug effects / Q000528:radiation effects; D002455:Cell Division / Q000187:drug effects / Q000528:radiation effects; D015687:Cell Hypoxia; D002478:Cells, Cultured; D002642:Chick Embryo; D003035:Cobalt / Q000494:pharmacology; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000235:genetics / Q000378:metabolism; D004050:Ditiocarb / Q000494:pharmacology; D005347:Fibroblasts; D005452:Fluoresceins / Q000032:analysis / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D051379:Mice; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D009206:Myocardium / Q000166:cytology; D017382:Reactive Oxygen Species / Q000378:metabolism*; D012402:Rotenone / Q000494:pharmacology; D013482:Superoxide Dismutase / Q000037:antagonists & inhibitors; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*; D014475:Uncoupling Agents / Q000494:pharmacology
chemical_list: D000975:Antioxidants; C493956:CDKN1A protein, human; C493957:Cdkn1a protein, mouse; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D005452:Fluoresceins; D017382:Reactive Oxygen Species; D016159:Tumor Suppressor Protein p53; D014475:Uncoupling Agents; D012402:Rotenone; D003035:Cobalt; C037631:2',7'-dichlorofluorescein; D004050:Ditiocarb; D006861:Hydrogen Peroxide; D013482:Superoxide Dismutase; C018021:cobaltous chloride; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid; D000111:Acetylcysteine