siRNA介导的STAT3基因沉默对宫颈癌HeLa细胞凋亡和化疗敏感性的影响

Effects of knockdown of STAT3 induced by small interference RNA on apoptosis of HeLa cells and chemoperapeutic sensitivity

目的:应用siRNA介导的STAT3基因沉默,研究其对HeLa细胞凋亡和化疗敏感性的影响。方法:体外合成靶向STAT3的siRNA-1和siRNA-2,并用脂质体转染HeLa细胞,RT-PCR检测干扰STAT3 mRNA敲减的效率,蛋白质印迹法检测敲减后STAT3蛋白的表达;通过记录生长曲线观察干扰后细胞的增殖情况。用Annexin V/PI双染,结合流式细胞仪检测细胞早期凋亡率。MTT法检测肿瘤细胞对几种化疗药物的敏感性。结果:siRNA使STAT3表达下调,mRNA水平抑制率为74.8%和60.4%;蛋白表达水平抑制达68.0%和59.0%。干扰后HeLa细胞增殖缓慢,早期凋亡率明显增加。对顺铂、长春瑞滨和吉西他滨有增敏作用。结论:siRNA介导的STAT3沉默可以抑制宫颈癌HeLa细胞的增殖,诱导凋亡,增加对化疗药物的敏感性,可望成为一种新的治疗策略。

OBJECTIVE：To investigate the apoptosis of HeLa cells induced by RNA interference （RNAi） targeting STAT3 and the effect on chemotherapeutic sensitivity. METHODS： Small interference RNA （siRNA-1 and -2） targeting STAT3 was generated by chemical synthesis and transfected into HeLa cells. The variance of STAT3 expressions was detected by semi-quantitive RT-PCR and Western blot. The proliferation and apoptosis of the transected cells were respectively detected by typan blue exclusion and FCM essay（Annexin V/PI）. MTT method was used to detect the chemoperapentic sensitvity. RESULTS： Both mRNA and protein levels of STAT3 in the HeLa cells were remarkably decreased after transfection with the two siRNAs,inhibited the reproduction and increased the apoptosis and the sensitivity to the chemotherapeutic drug. The mRNA inhibition rates were 74.8% and 60.4%. The protein inhibition rates were 68.0% and 59.0%. CONCLUSION： In vitro,STAT3 siRNA-1 and siRNA-2 transfection can specifically silence the gene expression and may induce apoptosis of HeLa cells,furthermore,elevate the sensitivity to chemotherapeutic drug.