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HepG2-1A2 C2 和 C7:慢病毒载体介导的人肝癌细胞细胞色素 P450 1A2 的稳定和功能性过表达。

HepG2-1A2 C2 and C7: Lentivirus vector-mediated stable and functional overexpression of cytochrome P450 1A2 in human hepatoblastoma cells.

机构信息

Institute of Biotechnology, Brandenburg University of Technology Cottbus-Senftenberg, Universitätsplatz 1, 01968 Senftenberg, Germany.

Project Group Pz-Syn of the Fraunhofer Institute for Cell Therapy and Immunology, Branch Bioanalytics and Bioprocesses IZI-BB, Am Mühlenberg 13, 14476 Potsdam, located at the Brandenburg University of Technology Cottbus-Senftenberg, Germany.

出版信息

Toxicol Lett. 2020 Feb 1;319:155-159. doi: 10.1016/j.toxlet.2019.11.006. Epub 2019 Nov 6.

Abstract

Novel HepG2 cell clones 1A2 C2 and 1A2 C7 were independently generated by lentiviral transduction to functionally overexpress cytochrome P450 1A2 (CYP1A2). We found similar and stable CYP1A2 transcript and protein levels in both cell clones leading to specific enzyme activities of about 370 pmol paracetamol x min x mg protein analyzed by phenacetin conversion. Both clones showed dramatically increased sensitivity to the hepatotoxic compound aflatoxin B (EC < 100 nM) when compared to parental HepG2 cells (EC5 μM). Thus, newly established cell lines are an appropriate tool to study metabolism and toxicity of substances depending on conversion by CYP1A2.

摘要

新型 HepG2 细胞克隆 1A2 C2 和 1A2 C7 是通过慢病毒转导独立生成的,以功能性过表达细胞色素 P450 1A2(CYP1A2)。我们发现这两个细胞克隆中的 CYP1A2 转录本和蛋白水平相似且稳定,导致通过苯乙酮转化分析的特定酶活性约为 370 pmol 对乙酰氨基酚 x min x mg 蛋白。与亲本 HepG2 细胞(EC5 μM)相比,这两个克隆对肝毒性化合物黄曲霉毒素 B(EC < 100 nM)的敏感性显著增加。因此,新建立的细胞系是研究依赖 CYP1A2 转化的物质代谢和毒性的合适工具。

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