Cytotoxicity and apoptosis produced by arachidonic acid in HepG2 cells overexpressing human cytochrome P-4502E1.
The goal of the current study was to evaluate the effects of arachidonic acid, as a representative polyunsaturated fatty acid, on the viability of a HepG2 cell line, which has been transduced to express human cytochrome P4502E1 (CYP2E1). Arachidonic acid produced a concentration- and time-dependent toxicity to HepG2-MV2E1-9 cells, which express CYP2E1, but little or no toxicity was found with control cells. In contrast to arachidonic acid, oleic acid was not toxic to the HepG2-MV2E1-9 cells. The cytotoxicity of arachidonic acid involved a lipid peroxidation type of mechanism since toxicity was enhanced after depletion of cellular glutathione; formation of malondialdehyde and 4-hydroxy-2-nonenal was markedly elevated in the cells expressing CYP2E1, and toxicity was prevented by antioxidants and the iron chelator desferrioxamine. The CYP2E1-dependent arachidonic acid toxicity appeared to involve apoptosis, as demonstrated by terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling and DNA laddering experiments. Trolox, which prevented toxicity of arachidonic acid, also prevented the apoptosis. Transfection with a plasmid containing bcl-2 resulted in complete protection against the CYP2E1-dependent arachidonic acid toxicity. It is proposed that elevated production of reactive oxygen intermediates by cells expressing CYP2E1 can cause lipid peroxidation, which subsequently promotes apoptosis and cell toxicity when the cells are enriched with polyunsaturated fatty acids such as arachidonic acid.
Chen
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Department of Biochemistry, Mount Sinai School of Medicine, New York, New York 10029, USA.
Galleano
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M
Cederbaum
A I
AI
D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics*; D016718:Arachidonic Acid / Q000494:pharmacology*; D002461:Cell Line, Transformed; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics*; D019392:Cytochrome P-450 CYP2E1 / Q000235:genetics*; D000431:Ethanol / Q000633:toxicity*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D006801:Humans; D015227:Lipid Peroxidation / Q000187:drug effects / Q000235:genetics*; D008114:Liver Neoplasms, Experimental; D014407:Tumor Cells, Cultured / Q000187:drug effects*
D016718:Arachidonic Acid; D000431:Ethanol; D019392:Cytochrome P-450 CYP2E1