干扰Yes相关蛋白表达调控Wnt/β-catenin信号通路影响膀胱癌细胞凋亡

Effect of interference in YAP expression on apoptosis of bladder cancer cells by regulation of Wnt/β-catenin signaling pathway

目的:探讨干扰Yes相关蛋白(YAP)的表达对膀胱癌细胞凋亡的影响及机制。方法:以膀胱癌细胞T-24为研究对象,分为正常对照(control)组、siRNA阴性对照(siRNA-NC)组和YAP siRNA组。分别以real-time PCR和Western blot实验检测转染后各组细胞中YAP的mRNA和蛋白表达水平,MTT法检测细胞活力,流式细胞术检测细胞凋亡情况,Western blot检测β-连环蛋白(β-catenin)和c-Myc表达水平。用Wnt/β-catenin信号通路抑制剂FH535处理YAP siRNA组细胞,并用MTT法检测细胞活力,流式细胞术检测细胞凋亡。结果:YAP siRNA组YAP的mRNA和蛋白水平均明显低于control组(P<0.05)。YAP siRNA组的细胞活力及c-Myc和β-catenin水平均明显低于control组(P<0.05),而细胞凋亡率明显高于control组(P<0.05)。与未经FH535处理的YAP siRNA组细胞相比,YAP siRNA组的细胞经FH535处理后细胞活力明显下降,细胞凋亡率升高,差异有统计学意义(P<0.05)。结论:干扰YAP表达能够通过抑制Wnt/β-catenin信号通路激活而抑制膀胱癌细胞活力,并促进膀胱癌细胞凋亡。

AIM： To investigate the effect of interference in Yes-associated protein（ YAP） expression on bladder cancer cell apoptosis and its mechanisms. METHODS： Bladder cancer cell line T-24 was used as the target and divided into control group（ no treatment）,small interfering RNA negative control（ siRNA-NC） group,and YAP siRNA group.The expression levels of YAP at mRNA and protein levels in the transfected cells of each group were detected by real-time PCR and Western blot,respectively. The cell viability was measured by MTT assay. The apoptosis was analyzed by flow cytometry. The protein expression levels of β-catenin and c-Myc were determined by Western blot. The cells in YAP siRNA group were treated with Wnt/β-catenin signaling pathway inhibitor FH535,and then the cell viability and apoptosis were analyzed by MTT assay and flow cytometry,respectively. RESULTS： The expression of YAP at mRNA and protein levels in YAP siRNA group was significantly lower than that in control group（ P 〈 0. 05）. The cell viability and the expression levels of c-Myc and β-catenin in YAP siRNA group were significantly lower than those in control group（ P 〈 0. 05）,while the apoptotic rate was significantly higher than that in control group（ P 〈 0. 05）. The cell viability in YAP siRNA ＋ FH535 group was decreased significantly,while the apoptotic rate was increased as compared with YAP siRNA group（ P 〈 0. 05）.CONCLUSION： Interfering with the expression of YAP inhibits the viability of bladder cancer cells by inhibiting the Wnt/β-catenin signaling pathway,thus promoting apoptosis of bladder cancer cells.