TY - JOUR
T1 - Stable expression of human cytochrome P450 2E1 in V79 Chinese hamster cells
AU - Schmalix, Wolfgang A.
AU - Barrenscheen, Martina
AU - Landsiedel, Robert
AU - Janzowski, Christine
AU - Eisenbrand, Gerhard
AU - Gonzalez, Frank
AU - Eliasson, Erik
AU - Ingelman-Sundberg, Magnus
AU - Perchermeier, Monika
AU - Greim, Helmut
AU - Doehmer, Johannes
PY - 1995/7/1
Y1 - 1995/7/1
N2 - 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.
AB - 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.
KW - Chlorzoxazone
KW - Cytochrome P450
KW - Cytochrome P450 2E1
KW - N-Nitrosodimethylamine
KW - p-Nitrophenol
KW - V79 Chinese hamster cell
UR - http://www.scopus.com/inward/record.url?scp=0028981769&partnerID=8YFLogxK
U2 - 10.1016/0926-6917(95)00008-9
DO - 10.1016/0926-6917(95)00008-9
M3 - Article
C2 - 7589226
AN - SCOPUS:0028981769
SN - 0926-6917
VL - 293
SP - 123
EP - 131
JO - European Journal of Pharmacology: Environmental Toxicology and
JF - European Journal of Pharmacology: Environmental Toxicology and
IS - 2
ER -