【摘要】 目的:构建Stat3-siRNA表达载体,探讨转染Stat3-siRNA表达载体阻断人胃癌细胞系SGC7901的Stat3信号传导通路对人胃癌细胞增殖和凋亡的影响。方法:利用RNAi技术,构建Stat3-siRNA表达载体,稳定转染胃癌细胞系SGC7901,利用Real-time PCR、Western Blot、MTT、流式细胞技术等方法分别检测Stat3基因mRNA和蛋白表达及细胞增殖和凋亡的变化情况。结果:成功构建Stat3-siRNA表达载体,经测序验证无误。Western Blot及Real-time PCR结果显示:Stat3基因的表达水平均下降,其中以SGC7901-siRNA3干扰效果最明显,差异具有显著性(P<0.05)。MTT法结果显示:稳定转染SGC7901细胞后,细胞的增殖能力明显下降,与对照组相比差别显著(P<0.05)。流式细胞技术显示:Stat3-siRNA3实验组较对照组出现了明显的细胞凋亡,在G1期前出现的亚二倍峰即为凋亡峰。结论:Stat3基因的RNAi重组体可以有效的抑制Stat3基因的表达,RNA干扰技术沉默Stat3基因能够抑制胃癌细胞的生长,促进其凋亡。更多还原

【Abstract】 Objective: To construct a small interfering RNA (siRNA) expression vector targeting the signal transducers and activators of transcription 3 (Stat3) gene and to investigate the impact and mechanism of a blockade of the stat3 signaling pathway in the SGC7901 cell line. Methods: After successful construction of a Stat3-siRNA expression vector, it was transfected into SGC7901. RT-PCR, Western blot, MTT and flow cytometry were used to detect the mRNA expression, the protein expression, cell proliferation and apoptosis. Results: A Stat3-siRNA expression vector was successfully constructed. The results of Western blot and RT-PCR showed that the expression of Stat3 decreased significantly (P<0.05), especially under the influence of siRNA3. MTT showed that cell proliferation was significantly reduced after transfection of the siRNA vector into SGC7901 cells. Flow cytometry results suggested that more apoptosis was seen in the siRNA group (P<0.05). Conclusion: RNAi of Stat3 can effectively inhibit Stat3 expression, suppress the proliferation of cancer cells, and promote apoptosis.更多还原