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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 effect of STAT3 gene silencing on HeLa cell apoptosis and chemosensitivity. Methods: siRNA-1 and siRNA-2 targeting STAT3 were synthesized in vitro and transfected into HeLa cells by lipofectamine. The efficiency of knockdown of STAT3 mRNA was detected by RT-PCR and the STAT3 protein expression was detected by Western blotting. Record growth curve to observe the proliferation of cells after interference. Annexin V / PI double staining, combined with flow cytometry cells early apoptosis rate. MTT assay of tumor cells on several chemotherapeutic drugs sensitivity. Results: siRNA down-regulated the expression of STAT3, the mRNA levels were inhibited by 74.8% and 60.4%, respectively. The protein expression levels were inhibited by 68.0% and 59.0%, respectively. HeLa cells proliferated slowly and the early apoptosis rate increased significantly after interference. Cisplatin, vinorelbine and gemcitabine sensitizing effect. Conclusion: siRNA mediated STAT3 silencing can inhibit the proliferation of cervical cancer HeLa cells, induce apoptosis and increase the sensitivity to chemotherapeutic drugs, which is expected to be a new therapeutic strategy.