Journal article
Increased prooxidant production and enhanced susceptibility to glutathione depletion in HepG2 cells co-expressing HCV core protein and CYP2E1
Journal of medical virology, Vol.72(2), pp.230-240
2004
DOI: 10.1002/jmv.10567
PMID: 14695664
Abstract
Hepatitis C virus (HCV) and HCV core protein are hypothesized to induce hepatic oxidative stress and exacerbate injury caused by other toxins such as ethanol that induce the cytochrome P450 enzyme, CYP2E1. In the current study, the effects of HCV core protein [sequence genotype 1b, (nt 342–915)] on parameters indicative of oxidative stress were evaluated in HepG2 cells stably over expressing CYP2E1 (E47), or vector controls (C34). Stable (>10 passages) expression of HCV core protein and CYP2E1 was confirmed in clonal cell lines at the level of mRNA and immunoreactive protein. Prooxidant production, as determined by cellular oxidation of dichlorodihydrofluorescin and dihydroethidium (HE), was increased by expression of HCV core protein in the presence or absence of CYP2E1. Depletion of glutathione (GSH) with buthionine sulfoximine (BSO) enhanced prooxidant production in both C34 and E47 cells. In addition, prooxidant production was greater in BSO‐treated cells expressing HCV core protein, and this effect was further enhanced in cells expressing both HCV core and CYP2E1. The CYP2E1 inhibitor, 4‐methylpyrazole, could suppress increased prooxidant production in E47 cells. Finally, cells co‐expressing both CYP2E1 and HCV core protein showed significantly decreased viability following GSH depletion. These studies show simultaneous expression of HCV core protein and CYP2E1 increases parameters indicative of oxidative stress as well as sensitization to cell injury induced by GSH depletion. These results support the hypothesis that enhanced injury in hepatocytes over expressing both HCV core protein and CYP2E1 is mediated by increases in oxidative stress.
Details
- Title: Subtitle
- Increased prooxidant production and enhanced susceptibility to glutathione depletion in HepG2 cells co-expressing HCV core protein and CYP2E1
- Creators
- Feng Wen - Department of Internal Medicine, University of Iowa College of Medicine, Iowa City, Iowa, United StatesMaher Y ABDALLA - Free Radical and Radiation Biology Program, Department of Radiation Oncology, University of Iowa, Iowa City, Iowa, United StatesBradley E BRITIGAN - Department of Internal Medicine, University of Iowa College of Medicine, Iowa City, Iowa, United StatesWarren N SCHMIDT - Department of Internal Medicine, University of Iowa College of Medicine, Iowa City, Iowa, United StatesCostica ALOMAN - Department of Internal Medicine, University of Iowa College of Medicine, Iowa City, Iowa, United StatesJinhua Xiang - Department of Internal Medicine, University of Iowa College of Medicine, Iowa City, Iowa, United StatesIman M AHMAD - Free Radical and Radiation Biology Program, Department of Radiation Oncology, University of Iowa, Iowa City, Iowa, United StatesJose WALEWSKI - Division of Liver Diseases, Department of Medicine, Mount Sinai School of Medicine, New York, New York, United StatesMichael L MCCORMICK - Department of Internal Medicine, University of Iowa College of Medicine, Iowa City, Iowa, United StatesKyle E BROWN - Department of Internal Medicine, University of Iowa College of Medicine, Iowa City, Iowa, United StatesAndrea D BRANCH - Division of Liver Diseases, Department of Medicine, Mount Sinai School of Medicine, New York, New York, United StatesDouglas R SPITZ - Free Radical and Radiation Biology Program, Department of Radiation Oncology, University of Iowa, Iowa City, Iowa, United States
- Resource Type
- Journal article
- Publication Details
- Journal of medical virology, Vol.72(2), pp.230-240
- Publisher
- Wiley-Liss; New York, NY
- DOI
- 10.1002/jmv.10567
- PMID
- 14695664
- ISSN
- 0146-6615
- eISSN
- 1096-9071
- Language
- English
- Date published
- 2004
- Academic Unit
- Gastroenterology and Hepatology; Pathology; Radiation Oncology; Internal Medicine
- Record Identifier
- 9984047706802771
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