Ethanol‐related cytotoxicity catalyzed by CYP2E1‐dependent generation of reactive oxygen intermediates in transduced HepG2 cells
- 1 January 1998
- journal article
- research article
- Published by Wiley in BioFactors
- Vol. 8 (1-2), 93-96
- https://doi.org/10.1002/biof.5520080116
Abstract
To establish direct linkage between the ethanol‐inducible cytochrome P450, CYP2E1, ethanol hepatotoxicity, and lipid peroxidation, a HepG2 cell line which expresses human CYP2E1 was established by retroviral infection. Ethanol produced a time‐ and concentration‐dependent cytotoxicity to HepG2 cells expressing the CYP2E1 but not to control cells. The ethanol toxicity was prevented by inhibitors of CYP2E1 and antioxidants. In a similar manner, addition of a polyunsaturated fatty acid such as arachidonic acid produced toxicity to the cells expressing CYP2E1 but not the control cells. Toxicity was associated with enhanced lipid peroxidation and was prevented by antioxidants. The ethanol and arachidonic acid toxicity was apoptotic in nature and was associated with activation of Caspases I and III. The toxicity and apoptosis could be prevented by peptide inhibitors of ICE and by transfection with a plasmid containing the cDNA for human Bcl‐2. These results show that this HepG2 cell model can be used to establish a CYP2E1‐dependent ethanol hepatotoxicity system, and that induction of a state oxidative stress appears to play a central role in the CYP2E1‐dependent apoptosis and cytotoxicity.Keywords
This publication has 10 references indexed in Scilit:
- Cytotoxicity and Apoptosis Produced by Arachidonic Acid in Hep G2 Cells Overexpressing Human Cytochrome P4502E1Journal of Biological Chemistry, 1997
- Ethanol Cytotoxicity to a Transfected HepG2 Cell Line Expressing Human Cytochrome P4502E1Journal of Biological Chemistry, 1996
- Ethanol Increases Content and Activity of Human Cytochrome P4502E1 in a Transduced HepG2 Cell LineBiochemical and Biophysical Research Communications, 1994
- Stable expression of human cytochrome P4502E1 in HepG2 cells: Characterization of catalytic activities and production of reactive oxygen intermediatesBiochemistry, 1993
- Hepatic Mitochondrial Glutathione Depletion and Progression of Experimental Alcoholic Liver Disease in RatsHepatology, 1992
- Implication of free radical mechanisms in ethanol-induced cellular injuryFree Radical Biology & Medicine, 1992
- Oxidative and reductive metabolism by cytochrome P450 2E1The FASEB Journal, 1992
- Role of human cytochrome P-450 IIE1 in the oxidation of many low molecular weight cancer suspectsChemical Research in Toxicology, 1991
- Insights into the pathogenesis of alcoholic liver necrosis and fibrosis: Status reportHepatology, 1990
- Oxygen radical generation by microsomes: Role of iron and implications for alcohol metabolism and toxicityFree Radical Biology & Medicine, 1989