RNAi沉默STAT3基因对人卵巢癌SKOV3细胞增殖和凋亡的作用

Effect of RNA Interference Targeting STAT3 Gene on Proliferation and Apoptosis of SKOV3 Cells

目的探讨信号转导与激活因子3(signal transduction and activators of transcription 3,STAT3)短发夹RNA(short hair-pin RNA,shRNA)真核表达载体对卵巢癌SKOV3细胞增殖和凋亡的作用。方法根据siRNA(small interference RNA,siR-NA)设计原则,结合pSilencer2.1-U6-neo质粒特点,针对STAT3基因设计并合成两条寡聚DNA片段,退火后克隆入pSilenc-er 2.1-U6-neo质粒,应用脂质体将重组质粒转染人卵巢癌SKOV3细胞。RT-PCR和Western blot检测SKOV3细胞STAT3基因mRNA和蛋白表达水平;MTT法和流式细胞仪检测细胞的增殖和凋亡。结果成功构建了STAT3基因shRNA真核表达载体,转染卵巢癌SKOV3细胞。细胞STAT3蛋白及mRNA表达下降,同时细胞增殖能力下降,凋亡率增加。结论构建的STAT3基因shRNA真核表达载体能够抑制卵巢癌细胞的增殖功能,诱导细胞凋亡。

[ Objective] To investigate the effect of eukaryotic vector expressing short hairpin RNA （shRNA） of signal transducers and activators of transcription 3 （ STAT3 ） on proliferation and apoptosis of SKOV3 cells. [ Methods ] According to siRNA design principles and characteristic of Psilencer2.1-U6-neo, 2 oligomeric DNA fragments were designed based on STAT3 gene sequence and was cloned into Psilencer2.1-U6-neo vector after annealing. The resultant plasmid was transfected into SKOV3 cells. The expressing levels of STAT3 gene mRNA and protein were detected by RT-PCR and Western blot. The cellular growth activity and apoptosis were assayed by MTT and flow cytometry. [ Results] Eukaryotic vector expressing shRNA of STAT3 was successfully constructed and transfected into SKOV3 cells. The expressions of STAT3 protein and mRNA in SKOV3 ceils decreased, meanwhile, the cellular growth activity decreased and cell apoptosis increased. [ Conclusion] The constructed shRNA eukaryotic expression vector of STAT3 can inhibit the cellular proliferation of SKOV3 and induce apoptosis.