【摘要】 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;②ERCC1 siRNA+顺铂组的ERCC1蛋白表达要弱于ERCC1 siRNA+PBS组,而GFP siRNA+顺铂组的蛋白表达要弱于GFP siRNA+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,ER-CC1siRNA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。更多还原

【Abstract】 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(MTT)colorimetric assay were used to determine the livability of the cancer cells in different Cisplatin concentration. Results:①After transfection of ERCC1 siRNA,The results showed the ERCC1 protein expression level of control group was significantly higher than that of experimental group;②ERCC1 siRNA+Cisplatin group and GFP siRNA+Cisplatin group expressed less ERCC1 protein than ERCC1 siRNA+PBS group and GFP siRNA+PBS group respectively;③Cisplatin(at the concentration of 0.75～12μmol/L)inhibited the livability of A2780 cells in a dose-dependent manner in despite of siRNA;④At 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.更多还原