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

Wolfgang A. Schmalix, Martina Barrenscheen, Robert Landsiedel, Christine Janzowski, Gerhard Eisenbrand, Frank Gonzalez, Erik Eliasson, Magnus Ingelman-Sundberg, Monika Perchermeier, Helmut Greim, Johannes Doehmer

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48 Scopus citations

Abstract

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.

Original languageEnglish
Pages (from-to)123-131
Number of pages9
JournalEuropean Journal of Pharmacology: Environmental Toxicology and
Volume293
Issue number2
DOIs
StatePublished - 1 Jul 1995
Externally publishedYes

Keywords

  • Chlorzoxazone
  • Cytochrome P450
  • Cytochrome P450 2E1
  • N-Nitrosodimethylamine
  • V79 Chinese hamster cell
  • p-Nitrophenol

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