TPX2基因在胃癌中的表达及其对胃癌细胞增殖凋亡的影响和机制研究

Expression of TPX2 gene in gastric cancer and its mechanism of proliferation and apoptosis in gastric cancer cells

目的探讨Xklp2靶蛋白(TPX2)基因在胃癌中的表达及其对胃癌细胞增殖凋亡的影响和机制。方法 RT-PCR及Western blotting检测胃癌组织及癌旁组织中TPX2 mRNA和蛋白表达;将NC-siRNA、TPX2-siRNA1、TPX2-siRNA2、TPX2-siRNA3转染人胃癌SGC-7901细胞,未转染任何siRNA作为对照组,Western blotting检测转染后各组细胞中TPX2蛋白表达;CCK8实验检测人胃癌SGC-7901细胞转染12、24、48 h后细胞增殖情况;细胞转染48 h后,流式细胞仪检测细胞凋亡;Western blotting检测PI3K、AKT、p-AKT蛋白表达。结果胃癌组织中TPX2 mRNA和蛋白表达均显著高于癌旁组织(P<0.01);NC-siRNA组TPX2蛋白表达与对照组比较,差异无统计学意义(P>0.05);TPX2-siRNA1、TPX2-siRNA2、TPX2-siRNA3组的TPX2蛋白表达均显著低于对照组(P<0.01),TPX2-siRNA3组的抑制效果更明显,选择作为后续研究;NC-siRNA组在12、24、48 h后的细胞存活率与对照组比较,差异无统计学意义(P>0.05),TPX2-siRNA组在12、24、48 h后的细胞存活率均显著低于对照组,且随着时间延长细胞存活率降低(P<0.01);NC-siRNA组细胞凋亡率及Cleaved caspase3、PI3K、AKT、p-AKT蛋白表达与对照组比较,差异无统计学意义(P>0.05),TPX2-siRNA组细胞凋亡率及Cleaved caspase3蛋白表达均显著高于对照组,PI3K、p-AKT蛋白表达显著低于对照组(P<0.01)。结论 TPX2在胃癌中高表达,沉默TPX2的表达能抑制人胃癌SGC-7901细胞增殖和促进细胞凋亡,其机制可能与PT3K/AKT信号通路的调控有关。

Objective To investigate the expression of TPX2 gene in gastric cancer and its effect on proliferation and apoptosis of gastric cancer cells. Methods The expressions of TPX2 mRNA and protein in gastric cancer tissues and adjacent tissues were detected by RT-PCR and Western blotting; NC-siRNA, TPX2-siRNA1, TPX2-siRNA2 and TPX2- siRNA3 were transfected into human gastric cancer SGC-7901 cells, and not transfected with any siRNA as the control group ; the expression of TPX2 protein in transfected cells was detected by Western blotting; cell proliferation transfected for 12, 24, 48 hours was detected by CCK8 assay; after cell transfection 48 h, cell apoptosis was detected by flow eytometry; the expressions of PI3K, AKT, p-AKT protein were detected by Western blotting. Results The expressions of TPX2 mRNA and protein in gastric cancer tissue were significantly higher than those in adjacent tissues （P 〈 0.01 ） ; the expression of TPX2 protein in NC-siRNA group and control group had no significant difference （P 〉 0.05） , the expression of TPX2 protein in TPX2-siRNAI, TPX2-siRNA2, TPX2-siRNA3 groups was significantly lower than that in the control group （P 〈0.01 ） , the inhibitory effect in TPX2-siRNA3 group was more obvious, was choiced as a follow- up study; the cell survival rate at 12, 24, 48 h in NC-siRNA group had no significant difference compared with control group （P 〉 0.05） , the cell survival rate at 12, 24, 48 h in TPX2-siRNA group was significantly lower than that in the control group, and the survival rate of cells was decreased with the time prolonging （P 〈 0.01 ） ; the apoptosis rate of cells and expressions of Cleaved caspase3, PI3K, AKT, p-AKT protein in NC-siRNA group had no significant difference compared with control group （P 〉 0.05） , the apoptosis rate of ceils and the expression of Cleaved caspase3 protein in TPX2-siRNA group were significantly higher than those in control group, the expressions of PI3K and p-AKT protein were significantly lower than those in the control group （P 〈 0.01 ）. Conclusion TPX2 is highly expressed in gastric cancer, the expression of TPX2 can inhibit the proliferation and apoptosis of human gastric cancer cell line SGC-7901, and its mechanism is related to the regulation of PT3K/AKT signaling pathway.