RNA干扰ERCC1基因表达对卵巢癌细胞耐药的影响

Effect on Cytotoxicity by RNAi ERCC1 Gene Expression in Ovarian Cancer CelI Line

目的:观察RNA干扰沉默ERCC1基因对卵巢癌细胞耐药的影响。方法:体外合成靶向于ERCC1基因的小干扰RNA(ERCC1-siRNA),用脂质体法转染至卵巢癌耐药细胞系COC1/DDP,RT-PCR方法检测转染前后细胞内ERCC1 mRNA的表达,W estern blot方法检测ERCC1基因蛋白表达的改变,MTT实验检测转染前后COC1/DDP细胞对顺铂敏感性的变化。结果:转染ERCC1-siRNA后,COC1/DDP细胞中ERCC1基因的mRNA和蛋白表达均明显减少。RNA干扰联合顺铂用药能明显提高COC1/DDP细胞对顺铂的敏感性。结论:RNA干扰ERCC1基因能明显抑制COC1/DDP细胞内ERCC1基因mRNA和蛋白的表达,增加COC1/DDP细胞对化疗药顺铂的敏感性。

Objective：To study the effect on drug resistance by RNAi mediated ERCC1 gene silencing in ovarian cancer cell line. Method：Small interference RNA targeted ERCC1 gene was synthesized in vitro and was transfected into COC1/DDP cell. The expression of ERCC1 was detected by semi - quantitative reverse transcription - polymerase chain reaction （ RT - PCR ） and western blot . MTT test were applied to measure the inhibition combined with cisplatin of COC1/DDP strain. Result： After transfection of ERCC1 - siRNA,mRNA and protein levels of ERCC1 gene in COC1/DDP cells were obviously reduced;ERCCl gene combined with cisplatin increases the sensitivity to chemotherapy. Conclusion： ERCC1 - siRNA could reduce the expression of ERCC1 gene and increase the sensitivity to cisplatin in COC1/DDP cell.