人细胞色素P4502E1cDNA在鼻咽癌细胞系CNE2的稳定表达.PDF

2001, 26( 6) BULL HUNAN MED UNIV 511 P450 2E1 cDNA CNE2 王水良, 贺智敏 , 陈主初 ( 中南大学湘雅医学院肿瘤研究所, 长沙 410078) [ ] 目的: 为进一步研究人细胞色素P450 E1( cytochrome P450 2E1, CYP2E1) 蛋白对相关化学致癌 的体外代 谢功能及其在化学 质鼻咽癌(naso haryngeal carcinoma, 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 CYP2E1 cDNA 真核表达载体 cDNA3. 1- 2E1, 并经脂质体介导转入CNE2 细胞, 通过G418 筛选后挑选若干 抗性细胞克隆扩大培养结果: 经Southern blot 和Western blot 对一系列抗性克隆细胞进行检测, 获得 2 个具有外源 CYP2E1 cDNA 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 筛选及CYP2E1 在NPC 癌变中作用机制研究的体外模型 [ ] 细胞色素P450 2E1; cDNA; 表达; 体外模型* [ ] R73963 [ ] A [ ] 2001) Stable Expression of Human Cytochrome P450 2E1 cDNA in Nasopharyngeal Cancer Cell Line CNE- 2 WANG Shuiliang, HE Zhimin , CHEN Zhuchu ( Cancer Research institute, X iangy a School of Medi cine , Central South University , Changsha 410078, China ) Abstract: Objecti e This study as designed to investigate the function of CYP2E1 rotein in chemical carcinogen metabolism in vitro and its action mechanisms in naso haryngeal carcinogenesis. Methods DAN re combination technique was used to construct a eukaryotic ex ression vector cDNA3. 1- 2E1 of human cy tochrome P450 2E1 cDNA. The vector was introduced into a naso haryngeal cancer cell line CNE- 2 mediating by li ofectamine. After these transforming cells were treated with G418 for 2 weeks, several G418- resistant cloneswere selected and am lified in culture. Results The authors acquired two cell clones CNE2- 2E1- 1 and CNE2- 2E1- 2 with stable integration and ex ression of exogenous CYPE2E1