Chen Zhongping, Zhang Junying, Mohr Gérard
Department of Neurosurgery, Cancer Center, Sun Yat-Sen University, Guangzhou 510060, China.
Chin Med J (Engl). 2003 Aug;116(8):1171-4.
To confirm the enhancing effect of excision repair cross complementing rodent repair deficiency gene 2 (ERCC2) on alkylating agents resistance.
The authors constructed a pcDNA3-ERCC2 plasmid. The pcDNA3-ERCC2 was transfected into a selected ERCC2 negative human glioma cell line, SKMG-4, using liposome-mediated transfection. After G418 selection, a stable transfected cell line was obtained and tested for cytotoxicity of several alkylating agents.
The stable transfectant was obtained and confirmed by RT-PCR as well as Western blot analysis to be strongly expressing ERCC2 at both mRNA and protein levels. The IC(90) ( micro mol/L) of two alkylating agents, cisplatin and melphalan, increased from 1.0 to 1.75 (75%) and 5.6 to 9.0 (61%), respectively, compared with control cell line.
The present data provided evidences and confirmed the authors' previous results that ERCC2 contributes, at least partially, to alkylating agent resistance in human glioma cell line.
证实切除修复交叉互补啮齿动物修复缺陷基因2（ERCC2）对烷化剂耐药性的增强作用。
构建pcDNA3-ERCC2质粒。采用脂质体介导转染法将pcDNA3-ERCC2转染入筛选出的ERCC2阴性人胶质瘤细胞系SKMG-4。经G418筛选后，获得稳定转染细胞系，并检测几种烷化剂的细胞毒性。
获得稳定转染子，经RT-PCR及蛋白质印迹分析证实其在mRNA和蛋白质水平均强烈表达ERCC2。与对照细胞系相比，顺铂和马法兰这两种烷化剂的IC(90)（微摩尔/升）分别从1.0增加到1.75（增加75%）和从5.6增加到9.0（增加61%）。
目前的数据提供了证据并证实了作者之前的结果，即ERCC2至少部分地导致人胶质瘤细胞系对烷化剂产生耐药性。
