抑制ERCC1基因对肺癌细胞顺铂敏感性的影响

RNA interference silencing excision repair cross-complementing 1 gene and affecting cisplatin sensitivity on lung cancer cell lines

目的探讨RNA干扰切除修复交叉互补基因（ERCC）1对肺腺癌细胞A549顺铂敏感性的影响。方法体外设计合成针对ERCC1基因的小干扰RNA（siRNA），转染肺腺癌细胞A549，应用RT—PCR、Western印迹和免疫细胞化学方法检测转染后ERCC1基因mRNA和蛋白的改变，应用四唑盐比色（MTT）法检测干扰ERCC1基因后A549细胞对顺铂敏感性的变化。结果转染针对ERCC1基因的siRNA后24、48、72h，A549细胞ERCC1基因mRNA和蛋白水平显著下降；关闭ERCC1基因后，A549细胞对顺铂的敏感性增加3．07倍。结论RNA干扰ERCC1基因能增加肺腺癌A549细胞对顺铂的敏感性。

Objective To research the changes of cisplatin sensitivity by RNA interfering the excision repair cross-complementing （ERCC） 1 gene in lung cancer cell lines. Methods The small interference RNA（siRNA） targeting ERCC1 gene was designed and synthesized by transcription in vitro, and transfected to lung cancer cell line A549. The mRNA and protein of ERCC1 were evaluated by means of RT- PCR, Western blot and immunocytochemistry. The changes of cisplatin sensitivity after interference were examined by methyl thiazolyl tetrazolium（MTT） assay. Results In A549 cell, the mRNA and protein levels of ERCC1 were dramatically decreased 24, 48 and 72 hours after transfeetion. The sensitivity to cisplatin of A549 cell line was increased by 3.07 times after disturbing the ERCC1 gene. Conclusions The sensitivity to eisplatin of lung cancer cell lines 3.549 could be enhanced by RNA interfering ERCC1 gene.