Wa. Schmalix et al., STABLE EXPRESSION OF HUMAN CYTOCHROME-P450 2E1 IN V79 CHINESE-HAMSTERCELLS, European journal of pharmacology. Environmental toxicology and pharmacology section, 293(2), 1995, pp. 123-131
A V79 Chinese hamster cell line was constructed for stable expression
of human cytochrome P450 2E1 (CYP2E1) by integration of a SV40 Early p
romoter recombinant CYP2E1 cDNA into the chromosomal DNA. The cDNA enc
oded CYP2E1 was effectively expressed and enzymatically active, as sho
wn by hydroxylation of chlorzoxazone and of p-nitrophenol, at rates of
about 70 pmol x mg(-1) total protein X min(-1). CYP2E1 content and ac
tivity was increased upon cultivation in the presence of ethanol indic
ating a substrate mediated stabilization effect. A similar stabilizing
effect was also observed for inhibitors of CYP2E1, e.g. imidazole, 4-
methylpyrazole, and isoniazid. The feasibility of the newly establishe
d cell line V79MZh2E1 for toxicological studies was shown by CYP2E1-me
diated activation of N-nitrosodimethylamine and p-nitrophenol and a do
se-dependent cytotoxic and mutagenic effect.