Cytochrome P-4502E1-dependent formation of trifluoroacetyl adducts from halothane by transduced HepG2 cells.
E-9 cells, a HepG2 cell line that has the alcohol-inducible human cytochrome P-4502E1 (CYP2E1) cloned into its genome, was tested for its ability to produce trifluoroacetyl (TFA) adducts from the metabolism of halothane. The metabolism of halothane results in the production of TFA halide that can readily react with cellular proteins to form TFA adducts, which can be detected by antibodies directed against them. E-9 cells formed TFA adducts when incubated with halothane. The major consistent bands that were detected were of molecular weights of approximately 50, 78, and 100 kDa. The formation of these adducts was dependent on halothane concentration and time of incubation with halothane. MV-5 cells, the control cell line that has the viral vector without the P-450, did not produce any adducts. Inhibitors of CYP2E1 function, such as 4-methylpyrazole, inhibited adduct formation. Furthermore, phorbol esters, which have been shown to increase the CYP2E1 level in this cell line, increased TFA adduct formation. This HepG2 cell line may be of value in studying the metabolism and toxicity of halothane in a human cell culture model.
Feierman
D E
DE
Department of Anesthesiology, Mount Sinai School of Medicine of the City University of New York, New York 10029-6574, USA.
Melnikov
Z
Z
D000818:Animals; D003001:Cloning, Molecular; D019392:Cytochrome P-450 CYP2E1 / Q000378:metabolism*; D004586:Electrophoresis; D000077604:Fomepizole; D006221:Halothane / Q000378:metabolism / Q000633:toxicity*; D006801:Humans; D008099:Liver / Q000378:metabolism; D010703:Phorbol Esters / Q000494:pharmacology; D011720:Pyrazoles / Q000494:pharmacology; D014269:Trifluoroacetic Acid / Q000378:metabolism; D014407:Tumor Cells, Cultured
D010703:Phorbol Esters; D011720:Pyrazoles; D000077604:Fomepizole; D014269:Trifluoroacetic Acid; D019392:Cytochrome P-450 CYP2E1; D006221:Halothane