过表达YAP基因通过PI3 K/AKT/mTOR信号通路促进舌鳞癌增殖

YAP gene overexpression promotes tongue squamous carcinoma growth through PI3K/AKT/mTOR signaling pathway

目的基于PI3K/AKT/mTOR信号通路,探究过表达YAP基因对舌鳞癌细胞增殖和凋亡的分子学调控机制.方法构建稳定过表达YAP基因的舌鳞癌CAL27和SCC25细胞系.采用CCK-8和流式细胞术分析过表达YAP基因对细胞增殖、周期和凋亡的影响.蛋白免疫印迹法分析过表达YAP基因对细胞内CDK4、CDK6、p21、BAX及对PI3K/AKT/mTOR信号通路的影响.结果CAL27细胞和SCC25细胞中过表达YAP组YAP mRNA较对照组升高(t=-19.29,P<0.001;t=-5.44,P<0.001),且过表达YAP组的YAP蛋白表达量较对照组增加(t=-31.28,P<0.001;t=-42.84,P<0.001);过表达YAP组的G1期细胞比例减少,过表达YAP组S期细胞比例较对照组升高;过表达YAP组的早期凋亡和晚期凋亡细胞都减少;过表达YAP组细胞中CDK4、CDK6、BCL-2、PARP等蛋白表达升高,p21、BAX及caspase3蛋白的表达减少;过表达YAP基因可使CAL27和SCC25细胞内AKT(tCAL27=-30.60,P<0.001;tSCC25=-16.77,P<0.001)、p-AKT(tCAL27=-10.23,P<0.001;tSCC25=-10.67,P<0.001),mTOR(tCAL27=-13.93,P<0.001;tSCC25=-9.57,P<0.001)、p-mTOR(tCAL27=-17.17,P<0.001;tSCC25=-8.51,P<0.001)表达量较对照组增高.结论过表达YAP基因可以通过PI3K/AKT/mTOR信号通路促进CAL27和SCC25细胞增殖并抑制细胞凋亡.

Objective To explore the molecular mechanism of YAP gene overexpression on the proliferation and apoptosis of tongue squamous carcinoma cells through the PI3K/AKT/mTOR signaling pathway.Methods CAL27 and SCC25 cell lines overexpressing YAP gene were constructed.The effects of YAP gene overexpression on cell proliferation,cell cycle and apoptosis were detected with CCK-8 and flow cytometry.Effects of YAP gene overexpression on the CDK4,CDK6,p21,BAX and PI3K/AKT/mTOR signaling pathways were analyzed with Western blotting.Results The mRNA expression of YAP in the overexpressing group was higher than that in the control group(t=-19.29,P<0.001;t=-5.44,P<0.001),and the protein expression was also higher(t=-31.28,P<0.001;t=-42.84,P<0.001).In the overexpressing group,the proportion of cells in G1 phase decreased,while the proportion of cells in S phase increased;the numbers of both early apoptosis and late apoptosis cells reduced;the protein expressions of CDK4,CDK6,BCL-2,PARP increased,while the protein expressions of p21,BAX and caspase3 decreased.YAP gene overexpression increased the expressions of AKT(tCAL27=-30.60,P<0.001;tSCC25=-16.77,P<0.001),p-AKT(tCAL27=-10.23,P<0.001;tSCC25=-10.67,P<0.001),mTOR(tCAL27=-13.93,P<0.001;tSCC25=-9.57,P<0.001),and pmTOR(tCAL27=-17.17,P<0.001;tSCC25=-8.51,P<0.001).Conclusion YAP gene overexpression promotes proliferation of CAL27 and SCC25 cells and inhibits apoptosis through the PI3K/AKT/mTOR signaling pathway.