背景与目的:NER是DNA修复系统中一个由多种蛋白组成的复杂网状系统,其中作为主要组成成分的ERCC1的表达同DDP化疗耐受相关,ERCC2的高表达同烷基药物化疗耐受相关。研究已经发现某些基因启动子区CpG岛的甲基化会导致基因的表达沉默,对化疗反应有指示作用,本文探讨ERCCI和ERCC2启动子区的甲基化状态和耐药的关系。方法:应用亚硫酸氢钠处理基因组DNA后PCR和不测序,我们分析了5个具有不同药敏特征的胶质瘤细胞株ERCC1和ERCC2的甲基化状态,人脑胶质瘤细胞株UW28,MGR1、MGR2、SF767和T98G和DDP和MeCCNU的敏感性采用MTT法检测。结果:在DDP敏感的细胞株中发现ERCC1基因启动子区上游4.9kb处存在甲基化的CpG岛,而在5个对BCNU耐药的细胞株中ERCC2的启动子区的CpG岛均呈去甲基化状态。结论:ERCC1和ERCC2畸变的甲基化状态与胶质瘤细胞株对化疗药的敏感性有关。
BACKGROUND & OBJECTIVE: NER pathway is a complex network of DNA-repair system which involved with many proteins. In this system, ERCCI over expression is known to he related with platinum-resistauce while ERCC2 over expression is related to BCNU-resistauce. It has heen proved that methylation of the CpG; island in the promoter region is associated with silencing of the genes. In this study we analysis the relationship of methylation status in the ERCC1 and ERCC2 promoter and auti-vaneer drug resistance in glioma cell lines. METHODS: Using bisulphate sequeneing, we have identified hypennethylation in the promoter region of ERCC1 and ERCC2 in five gliomas cell lines.UW28、MGR1、MGR2、SF767 and T98G.The sensitivity of the cell lines to anti-cancer drug MeCCNU and DDP were determined by MTT assay.RESULTS:The methylated CpG is land in the 4.9kb region of ERCC1 gene promoter upstream in the platinum-sensitive cell lines has been recognized and the putative CpG island of ERCC2 promoter showed the ummethylated status in all five BCNU-resistant cell lines.CONCLUSION:Our data indicate that aberrant methylation status of ERCC1 and ERCC2 were associated with chemoensitivity in gliomas cell lines.