Gab2基因在胃癌中的表达及对胃癌细胞增殖、凋亡、迁移及AKT、p-AKT、Bax、Bcl-2表达的影响研究

目的 探讨Gab2基因在胃癌中的表达及对胃癌细胞增殖、凋亡、迁移及对AKT、p-AKT、Bax、Bcl-2表达的影响.方法 采用RT-PCR及Western Blot检测胃癌组织和癌旁组织中Gab2的表达.培养人胃癌细胞株SGC-7901,分别用Gab2小干扰RNA(Gab2-siRNA)和阴性对照(siRNA-NC)转染细胞,以空脂质体转染的细胞作为对照组,各组细胞培养48 h.应用Western Blot检测细胞中Gab2、AKT、p-AKT、Bax、Bcl-2蛋白表达的变化,CCK-8检测细胞增殖情况,流式细胞仪检测细胞凋亡,Transwell小室检测细胞迁移能力.结果 胃癌组织中Gab2的mRNA及蛋白表达水平均明显高于癌旁组织(P﹤0.01);Gab2-siRNA组Gab2蛋白表达水平明显低于对照组(P﹤0.01);siRNA-NC组细胞的存活率、凋亡率、迁移数及AKT、p-AKT、Bax、Bcl-2蛋白表达与对照组比较,差异均无统计学意义(P﹥0.05);Gab2-siRNA组细胞的AKT蛋白表达与对照组比较,差异无统计学意义(P﹥0.05),细胞的存活率、迁移数及p-AKT、Bcl-2蛋白表达明显低于对照组(P﹤0.01),细胞凋亡率及Bax蛋白表达明显高于对照组(P﹤0.01).结论 Gab2在胃癌组织高表达,沉默Gab2的表达能显著抑制人胃癌细胞株SGC-7901的增殖和迁移,并通过调节Bax、Bcl-2蛋白表达促进细胞凋亡,其可能的机制与AKT信号通路的调控有关.

The expression of Gab2 gene in gastric cancer and its effect on proliferation,apoptosis, and migration of cells and expression of AKT,p-AKT,Bax and Bcl-2 protein

Objective To investigate the expression of Gab2 gene in gastric cancer and its effect on the proliferation, apoptosis, migration of cancer cells and expression of AKT, p-AKT, Bax and Bcl-2 protein. Method The expression of Gab2 was detected in gastric cancer tissues and adjacent tissues by RT-PCR and Western Blot;Human gastric cancer cell line SGC-7901 were cultured, Gab2 small interfering RNA (Gab2- siRNA) and negative control (siRNA-NC) were re-spectively transfected into test cell line, while cells transfected with blank liposomes were used as the control group, all cells were cultured for 48 h;and then the expression of Gab2, AKT, p-AKT, Bax, Bcl-2 protein were detected by Western Blot;cell proliferation by CCK-8 assay, and cell apoptosis by flow cytometry;Transwell was used to evaluate cell migra-tion. Result The mRNA and protein expression of Gab2 in gastric cancer tissues were significantly higher than those in normal adjacent tissues (P<0.01);the expression level of Gab2 protein in Gab2-siRNA group was significantly lower than that of the control group (P<0.01);the survival rate, apoptosis rate, cell migration rate and AKT, p-AKT, Bax, Bcl-2 pro-tein expression in siRNA-NC group were similar with those of the control group (P>0.05);and so did the expression of AKT protein (P>0.05), while the survival rate, cell migration rate and p-AKT, Bcl-2 protein expression were significantly lower (P<0.01), and the apoptosis rate and the expression of Bax protein was significantly higher than those of the control group (P<0.01). Conclusion Gab2 is highly expressed in gastric cancer tissues;the proliferation and migration of human gastric cancer cell line SGC-7901 will be significantly inhibited as Gab2 is silenced, and can promote cell apoptosis by regulating the expression of Bax, Bcl-2 protein, of which the possible mechanism is associated with the regulation of AKT signaling pathway.