Stable expression of human cytochrome P450 2E1 in V79 Chinese hamster cells

A V79 Chinese hamster cell line was constructed for stable expression of human cytochrome P450 2E1 (CYP2E1) by integration of a SV40 Early promoter recombinant CYP2E1 cDNA into the chromosomal DNA. The cDNA encoded CYP2E1 was effectively expressed and enzymatically active, as shown by hydroxylation...

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Published in:European journal of pharmacology. Environmental toxicology and pharmacology section 1995-07, Vol.293 (2), p.123-131
Main Authors: Schmalix, Wolfgang A., Barrenscheen, Martina, Landsiedel, Robert, Janzowski, Christine, Eisenbrand, Gerhard, Gonzalez, Frank, Eliasson, Erik, Ingelman-Sundberg, Magnus, Perchermeier, Monika, Greim, Helmut, Doehmer, Johannes
Format: Article
Language:English
Subjects:
Chlorzoxazone
Cytochrome P450
Cytochrome P450 2E1
N-Nitrosodimethylamine
p-Nitrophenol
V79 Chinese hamster cell
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Summary:A V79 Chinese hamster cell line was constructed for stable expression of human cytochrome P450 2E1 (CYP2E1) by integration of a SV40 Early promoter recombinant CYP2E1 cDNA into the chromosomal DNA. The cDNA encoded CYP2E1 was effectively expressed and enzymatically active, as shown by hydroxylation of chlorzoxazone and of p-nitrophenol, at rates of about 70 pmol × mg −1 total protein × min −1. CYP2E1 content and activity was increased upon cultivation in the presence of ethanol indicating 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 established cell line V79MZh2E1 for toxicological studies was shown by CYP2E1-mediated activation of N-nitrosodimethylamine and p-nitrophenol and a dose-dependent cytotoxic and mutagenic effect.
ISSN:0926-6917
DOI:10.1016/0926-6917(95)00008-9