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作 者:韩建秋[1] 吴维光[2] 葛红雨[2] 王勇梅[2]
机构地区:[1]华北煤炭医学院研究生部,唐山063000 [2]武警医学院附属医院妇产科,天津300162
出 处:《重庆医科大学学报》2010年第10期1506-1509,共4页Journal of Chongqing Medical University
摘 要: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.
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