RNAi沉默STAT3基因对人卵巢癌SKOV3细胞增殖和凋亡的影响

Effects of RNA interference targeting STAT3 gene on proliferation and apoptosis of SKOV3 cells

目的:探讨应用RNAi技术沉默信号转导与激活因子3(Signal transducers and activators of transcription 3,STAT3)基因对人卵巢癌SKOV3细胞的影响。方法:根据siRNA(Small interference RNAs,iRNA)设计原则,结合pSilencer2.1-U6-neo质粒特点,针对STAT3基因设计并合成2条寡聚DNA片段,退火后克隆入pSilencer2.1-U6-neo质粒,应用脂质体将重组质粒转染SKOV3细胞。通过RT-PCR及Western blot检测STAT3基因不同水平的表达,采用MTT实验、流式细胞仪检测细胞增殖、凋亡的变化。结果:目的siRNA实验组细胞STAT3蛋白及mRNA表达水平均明显下降,细胞增殖能力显著降低,而细胞凋亡率则显著升高。结论:应用RNAi技术沉默STAT3基因可以降低卵巢癌SKOV3细胞STAT3的表达,进而抑制细胞的生长、增殖及诱导细胞的凋亡。

Objective： To investigate the inhibition ofSKOV3 cell by applying RNAi technique to silence signal transducers and activators of transcription 3 （STAT3）gene. Methods： shRNA templates was designed on the basis of STAT3 gene sequence and was cloned into pSilencer2.1-U6-neo vector. The resultant plasmid was transfected into SKOV3 cells with Lipofectamine 2000. The STAT3 protein and mRNA were detected by Western blot and RT-PCR,respectively. The cellular growth activity was assayed by MTT,and the apoptosis was tested by flow cytometry. Results： STAT3 protein and mRNA expression in target siRNA group were decreased ,respectively. In target siRNA group,cell proliferation was inhibited obviously and apoptosis rate increased significantly. Conclusion： Silencing STAT3 gene by the RNAi technology can decrease the expression of STAT3 gene,suppress the growth and proliferation of SKOV3 cell,and induce apoptosis of SKOV3 cell.