title: Stable expression of human cytochrome P450 2E1 in V79 Chinese hamster cells.
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 x mg-1 total protein x 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.
authors.affiliation: Institut für Toxikologie und Umwelthygiene, Technische Universität München, Germany.
authors.forename: W A
authors.initials: WA
authors.lastname: Schmalix
authors.forename: M
authors.initials: M
authors.lastname: Barrenscheen
authors.forename: R
authors.initials: R
authors.lastname: Landsiedel
authors.forename: C
authors.initials: C
authors.lastname: Janzowski
authors.forename: G
authors.initials: G
authors.lastname: Eisenbrand
authors.forename: F
authors.initials: F
authors.lastname: Gonzalez
authors.forename: E
authors.initials: E
authors.lastname: Eliasson
authors.forename: M
authors.initials: M
authors.lastname: Ingelman-Sundberg
authors.forename: M
authors.initials: M
authors.lastname: Perchermeier
authors.forename: H
authors.initials: H
authors.lastname: Greim
mesh_terms: D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D015153:Blotting, Western; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002753:Chlorzoxazone / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D019392:Cytochrome P-450 CYP2E1; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis* / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D008861:Microsomes / Q000187:drug effects / Q000201:enzymology; D009153:Mutagens / Q000633:toxicity; D009596:Nitrophenols / Q000378:metabolism; D010089:Oxidoreductases, N-Demethylating / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis; D014162:Transfection
chemical_list: D065607:Cytochrome P-450 Enzyme Inhibitors; D009153:Mutagens; D009596:Nitrophenols; D011994:Recombinant Proteins; D003577:Cytochrome P-450 Enzyme System; D019392:Cytochrome P-450 CYP2E1; D010089:Oxidoreductases, N-Demethylating; D002753:Chlorzoxazone; C024836:4-nitrophenol