NOB1基因对膀胱癌细胞凋亡的影响及其机制研究

目的探讨干扰NOB1基因对膀胱癌细胞凋亡和Wnt/β-catenin信号通路的影响。方法将膀胱癌T24细胞分为对照组(CON组),不进行转染、siRNA阴性对照组(siRNA-CON)、NOB1-siRNA组。分别采用RT-PCR和Western blot检测转染后三组细胞中NOB1 mRNA和蛋白的表达水平;MTT法检测细胞活力;流式细胞技术检测细胞凋亡情况;Western blot检测β-catenin和c-Myc蛋白表达水平。用Wnt/β-catenin信号通路抑制剂FH535处理NOB1-siRNA组细胞,设置为NOB1-siRNA-FH535组,检测细胞活力、细胞凋亡情况和β-catenin、c-Myc蛋白表达水平。结果NOB1-siRNA组NOB1 mRNA和蛋白表达水平均低于siRNA-CON组和CON组,差异有统计学意义(P<0.05)。NOB1-siRNA组细胞活力低于siRNA-CON组和CON组,差异有统计学意义(P<0.05)。流式细胞检测结果显示,NOB1-siRNA组细胞凋亡率高于siRNA-CON组和CON组,差异有统计学意义(P<0.05)。NOB1-siRNA组β-catenin和c-Myc蛋白表达水平均低于siRNA-CON组和CON组,差异有统计学意义(P<0.05)。NOB1-siRNA-FH535组细胞活力低于NOB1-siRNA组(P<0.05)。流式细胞技术检测结果显示,NOB1-siRNA-FH535组细胞凋亡率高于NOB1-siRNA组(P<0.05)。Western blot检测结果显示,NOB1-siRNA-FH535组β-catenin及c-Myc蛋白表达水平低于NOB1-siRNA组(P<0.05)。结论干扰NOB1表达可能通过抑制Wnt/β-catenin信号通路的激活而促进膀胱癌细胞的凋亡。

Effect of NOB1 gene on apoptosis of bladder cancer cell and its mechanism

Objective To investigate the effect of NOB1 gene interference on apoptosis and Wnt/beta-catenin signaling pathway in bladder cancer cells.Methods Bladder cancer T-24 cells were divided into control group(CON group),without transfection,siRNA-CON negative control group(siRNA-CON)and NOB1-siRNA group.The expression levels of NOB1 mRNA and protein were detected by RT-PCR and Western blot,cell viability was detected by MTT,cell apoptosis was detected by flow cytometry,and the expression levels of beta-catenin and c-Myc protein were detected by Western blot.NOB1-siRNA cells were treated with Wnt/β-catenin signaling pathway inhibitor FH535.The cells were divided into NOB1-siRNA-FH535 group.Cell viability,apoptosis and expression of beta-catenin and c-Myc protein were detected.Results The level of NOB1 gene and protein expression in NOB1-siRNA group was lower than that in siRNA-CON group and CON group,and the difference was significant(P<0.05).The cell viability of NOB1-siRNA group was lower than that of siRNA-CON group and CON group,the difference was significant(P<0.05).The results of flow cytometry showed that the apoptotic rate of NOB1-siRNA group was higher than that of siRNA-CON group and CON group(P<0.05).The expression levels of beta-catenin and c-Myc protein in NOB1-siRNA group were lower than those in siRNA-CON group and CON group(P<0.05).The cell viability of NOB1-siRNA-FH535 group was lower than that of NOB1-siRNA group(P<0.05).Flow cytometry showed that the apoptotic rate of NOB1-siRNA-FH535 group was higher than that of NOB1-siRNA group(P<0.05).Conclusions Interference of NOB1 expression may promote apoptosis of bladder cancer cells by inhibiting the activation of Wnt/β-catenin signaling pathway.