人细胞色素P450 2E1 cDNA在鼻咽癌细胞系CNE-2的稳定表达

目的 :为进一步研究人细胞色素P45 0E1 (cytochromeP45 0 2E1 ,CYP2E1 )蛋白对相关化学致癌物的体外代谢功能及其在化学物质鼻咽癌 (nasopharyngealcarcinoma ,NPC)变中的作用机制提供体外模型。方法 :应用基因重组技术构建人CYP2E1cDNA真核表达载体pcDNA3 .1 - 2E1 ,并经脂质体介导转入CNE2细胞 ,通过G41 8筛选后挑选若干抗性细胞克隆扩大培养。结果 :经Southernblot和Westernblot对一系列抗性克隆细胞进行检测 ,获得 2个具有外源CYP2E1cDNA基因稳定整合和表达的细胞克隆CNE2 - 2E1 - 1及CNE2 - 2E1 - 2。结论 :建立了可供CYP2E1代谢相关的化学致癌物筛选及CYP2E1在NPC癌变中作用机制研究的体外模型。

Stable Expression of Human Cytochrome P450 2E1 cDNA in Nasopharyngeal Cancer Cell Line CNE-2

Objective This study as designed to investigate the function of CYP2E1 protein in chemical carcinogen metabolism in vitro and its action mechanisms in nasopharyngeal carcinogenesis. Methods DAN recombination technique was used to construct a eukaryotic expression vector pcDNA3.1-2E1 of human cytochrome P450 2E1 cDNA. The vector was introduced into a nasopharyngeal cancer cell line CNE-2 mediating by lipofectamine. After these transforming cells were treated with G418 for 2 weeks, several G418-resistant clones were selected and amplified in culture. Results The authors acquired two cell clones CNE2-2E1-1 and CNE2-2E1-2 with stable integration and expression of exogenous CYPE2E1 cDNA. After Southern blot analysis, RT-PCR and Western blot were fulfilled. Conclusion The cell models setting here will be a good tool in the further study of CYP2E1. At the same time, its use will help to screen chemical carcinogenesis and to elucidate the mechanism of carcinogenesis which is related to chemical carcinogen activated by CYP2E1.