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[目的]探讨siRNA介导抗增殖蛋白prohibitin(PHB)基因沉默对胃癌细胞增殖凋亡的影响及机制。[方法]Western blot法检测胃癌组织及相应的癌旁组织中prohibitin的蛋白表达;NC-siRNA和prohibitin-siRNA转染人胃癌SGC-7901细胞,未转染任何的siRNA作为空白对照组,48h后检测各组细胞中prohibitin的蛋白表达;CCK8实验检测细胞增殖;流式细胞术检测细胞凋亡;Western blot检测Ki67、PCNA、Cleaved caspase3、PI3K、AKT、p-AKT蛋白表达。[结果]prohibitin在胃癌组织中的蛋白表达显著高于癌旁组织(P<0.01),转染prohibitin-siRNA后能显著降低prohibitin的蛋白表达;与对照组及NC-siRNA组比较,prohibitin-siRNA组细胞存活率显著降低,细胞凋亡率显著升高,Cleaved caspase3蛋白表达显著上调,ki67、PCNA、PI3K、p-AKT蛋白表达显著下调(P<0.01)。[结论]靶向prohibitin-siRNA干扰可有效的降低胃癌SGC-7901细胞中prohibitin的表达,抑制细胞增殖和诱导细胞凋亡,其机制与PI3K/AKT信号通路的调控有关。
[Objective] To investigate the effect of siRNA-mediated inhibition of anti-proliferative protein prohibitin (PHB) on proliferation and apoptosis of gastric cancer cells and its mechanism. [Methods] Western blot was used to detect the expression of prohibitin protein in gastric cancer tissues and its corresponding paracancerous tissues. NC-siRNA and prohibitin-siRNA transfected SGC-7901 cells without any siRNA were used as blank control group. After 48h The expression of prohibitin in each group of cells was detected; the proliferation of cells was detected by CCK8 assay; the apoptosis was detected by flow cytometry; the protein expression of Ki67, PCNA, Cleaved caspase3, PI3K, AKT and p-AKT were detected by Western blot. [Results] The expression of prohibitin protein in gastric cancer tissues was significantly higher than that in adjacent non-cancerous tissues (P <0.01). Prohibitin-siRNA significantly decreased the expression of prohibitin after transfection with prohibitin-siRNA. Compared with the control and NC- The cell viability was significantly decreased and the apoptosis rate was significantly increased. The expression of Cleaved caspase3 was significantly up-regulated, and the expressions of ki67, PCNA, PI3K and p-AKT were significantly down-regulated (P <0.01). [Conclusion] Targeting prohibitin-siRNA interference can effectively reduce prohibitin expression, inhibit cell proliferation and induce apoptosis in gastric cancer SGC-7901 cells. The mechanism is related to the regulation of PI3K / AKT signaling pathway.