Wolfgang A. Schmalix , Martina Barrenscheen , Robert Landsiedel , Christine Janzowski , Gerhard Eisenbrand , Frank Gonzalez , Erik Eliasson , Magnus Ingelman-Sundberg , Monika Perchermeier , Helmut Greim , Johannes Doehmer
{"title":"人细胞色素P450 2E1在中国仓鼠V79细胞中的稳定表达","authors":"Wolfgang A. Schmalix , Martina Barrenscheen , Robert Landsiedel , Christine Janzowski , Gerhard Eisenbrand , Frank Gonzalez , Erik Eliasson , Magnus Ingelman-Sundberg , Monika Perchermeier , Helmut Greim , Johannes Doehmer","doi":"10.1016/0926-6917(95)00008-9","DOIUrl":null,"url":null,"abstract":"<div><p>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 <em>p</em>-nitrophenol, at rates of about 70 pmol × mg<sup>−1</sup> total protein × min<sup>−1</sup>. 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 <em>N</em>-nitrosodimethylamine and <em>p</em>-nitrophenol and a dose-dependent cytotoxic and mutagenic effect.</p></div>","PeriodicalId":100501,"journal":{"name":"European Journal of Pharmacology: Environmental Toxicology and Pharmacology","volume":"293 2","pages":"Pages 123-131"},"PeriodicalIF":0.0000,"publicationDate":"1995-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0926-6917(95)00008-9","citationCount":"43","resultStr":"{\"title\":\"Stable expression of human cytochrome P450 2E1 in V79 Chinese hamster cells\",\"authors\":\"Wolfgang A. Schmalix , Martina Barrenscheen , Robert Landsiedel , Christine Janzowski , Gerhard Eisenbrand , Frank Gonzalez , Erik Eliasson , Magnus Ingelman-Sundberg , Monika Perchermeier , Helmut Greim , Johannes Doehmer\",\"doi\":\"10.1016/0926-6917(95)00008-9\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>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 <em>p</em>-nitrophenol, at rates of about 70 pmol × mg<sup>−1</sup> total protein × min<sup>−1</sup>. 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 <em>N</em>-nitrosodimethylamine and <em>p</em>-nitrophenol and a dose-dependent cytotoxic and mutagenic effect.</p></div>\",\"PeriodicalId\":100501,\"journal\":{\"name\":\"European Journal of Pharmacology: Environmental Toxicology and Pharmacology\",\"volume\":\"293 2\",\"pages\":\"Pages 123-131\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1995-07-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/0926-6917(95)00008-9\",\"citationCount\":\"43\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"European Journal of Pharmacology: Environmental Toxicology and Pharmacology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/0926691795000089\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"European Journal of Pharmacology: Environmental Toxicology and Pharmacology","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/0926691795000089","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
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 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.
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