RNA干扰抑制ERCC1基因对肺癌细胞A549药物敏感性影响的研究

Effects of inhibition of ERCC1 expression by RNA interference on chemotherapy sensitivity of lung cancer A549 cell line

目的:用RNA干扰技术下调肺癌细胞系A549切除修复交叉互补基因(ER-CC1)的表达,以提高肺癌细胞对化学药物的敏感性。方法:首先构建针对ERCC1基因的SiRNA真核表达载体;其次将载体转染肺癌细胞A549,采用RT-PCR和蛋白质印迹法技术证实转染细胞中ERCC1基因沉默效果良好;最后采用MTT法检测构建真核表达载体转染细胞对顺铂药物毒副反应的影响。结果:成功构建针对ERCC1基因的siRNA真核表达载体;将其转染肺癌细胞A549,可以抑制细胞中ERCC1基因表达,转染细胞较亲本细胞ERCC1基因表达降低48.3%,P<0.05;RNA干扰的A549细胞较亲本细胞对顺铂的敏感性升高了2.93倍。结论:RNA干扰是一种沉默ER-CC1表达的理想方法。沉默ERCC1表达可以提高肺癌细胞的化疗敏感性,它可能是一种有潜质的肺癌辅助治疗新方法。

OBJECTIVE： To study the inhibition of ERCC1 expression by RNA interference to improve chemotherapy sensitiv ity of lung cancer A549 cell line. METHODS：The small interference RNA （siRNA） of ERCC1 expression vector was established. The established vectors were transfect into lung cancer cells （A549） by the liposome transfection reagent. RT PCR and Western blotting were used to confirm whether the siRNA silenced the expression of ERCC1 in A549 cells. The characteristics of drug resistance in transfection cells were measured by MTT assay. RESULTS： siRNA of ERCC1 expression vector was constructed successfully. RT-PCR and Western blotting assay showed that the expression of ERCC1 in transfeeted A549 cells was significantly lower than that inA549 cells by 48.3%, P〈0.05. The values of IC50 of A549 cells to chemotherapy drugs（DDP） were 2.93 times higher than A549/vecter cells. CONCLUSIONS： The RNA interference to ER- CC1 gene is a good method to silence expression of ERCC1 in lung cancer cells. Inhibiting the expression of ERCC1 can enhance sen sitivity to chemotherapy drugs in lung cancer cells. It may be an available therapeutic method in the treatment of lung cancer.