沉默ERCC1基因对卵巢癌细胞A2780顺铂耐药性的影响

Impacts of ERCC1 silencing on Cisplatin resistance in ovarian cancer cell

目的：研究RNA干扰切除修复交叉互补基因1（ERCC1）的表达后，卵巢癌细胞A2780对顺铂耐药性的变化。方法：A2780细胞分别转染ERCC1基因的siRNA和GFPsiRNA，并给予顺铂（使其终浓度达到3μmol／L），蛋白质印迹法分别检测顺铂给药前后ERCC1蛋白的表达，并用MTT法检测顺铂不同浓度下A2780细胞的存活率。结果：①转染ERCC1 siRNA后，实验组蛋白质表达量较对照组显著降低；@ERCC1 siRNA＋顺铂组的ERCC1蛋白表达要弱于ERCC1 siRNA＋PBS组，而GFPsiRNA＋顺铂组的蛋白表达要弱于GFPsiRNA＋PBS组；③顺铂（浓度在0．75—12μmol／L之间）呈浓度依赖性地降低A2780细胞的存活率；④在相同浓度顺铂的作用下，与GFPsiRNA组相比，ERCC1 siRNA组的顺铂量效曲线左移。结论：顺铂能抑制沉默ERCC1基因后A2780细胞的生长，且在一定浓度范围内呈浓度依赖性；ERCC1是参与顺铂耐药性产生的重要因素之一。

Objective： To investigate the influence of RNA interfering the expression of excision repair cross-complementing 1 （ERCC1） gene on the sensitivity of ovarian cancer cell A2780 to Cisplatin. Methods： SiRNA and GFP siRNA was transfected to knock out ERCC1 gene instantaneous and make a control manner respectively. Cisplatin（ final concentration of 3μmol/L） was given to the knockouted cells. Western blot was taken for the analyses of ERCC1 protein expression in A2780 cells before and after Cisplatin was given. Methyl- thiazolyl tetrazolium （M337） colorimetric assay were used to determine the livability of the cancer cells in different Cisplatin concentration. Results： （1）After transfection of ERCC1 siRNA,The results showed the ERCC1 protein expression level of control group was significantly higher than that of experimental group; （2）ERCC1 siRNA ＋ Cisplatin group and GFP siRNA ＋ Cisplatin group expressed less ERCC1 protein than ERCC1 siRNA ＋ PBS group and GFP siRNA ＋ PBS group respectively;（3）Cisplatin （at the concentration of 0.75 - 12μmol/L） inhibited the livability of A2780 cells in a dose-dependent manner in despite of siRNA; （4）At a same concentration of Cisplatin, the dose-effect curve of knockout group was left to that of control group. Conclusion ： Cisplatin inhibited the proliferation of A2780 cells in a dose-dependent manner in despite of siRNA; ERCC1 is one of the important reasons of Cisplatin resistance in A2780 cell.