UNC5A基因过表达对膀胱癌T24细胞增殖、转移的影响及其机制探讨

Effects of UNC5A Gene Overexpression on Proliferation and Metastasis of Bladder Cancer T24 Cells and Its Mechanism

目的探究UNC5A基因过表达对膀胱癌T24细胞增殖、转移及Wnt/β-catenin信号通路的影响。方法选取T24细胞,转染UNC5A-pcDNA和pcDNA-NC,将细胞随机分为Control组、pcDNA-NC组和UNC5A-pcDNA组。RT-PCR检测UNC5A mRNA表达水平;克隆形成实验检测细胞克隆形成率;流式细胞术检测细胞凋亡率;Transwell检测细胞侵袭细胞数;Western印迹检测UNC5A、Wnt1、β-catenin蛋白表达水平;加入Wnt/β-catenin信号通路激活剂LiCl检测各组细胞增殖、凋亡、侵袭情况。结果与Control组比较,UNC5A-pcDNA组UNC5A mRNA和蛋白表达水平升高(P<0.05),克隆形成率降低(P<0.05),细胞凋亡率升高(P<0.05),侵袭细胞数降低(P<0.05),Wnt1、β-catenin蛋白表达水平降低(P<0.05)。加入Wnt/β-catenin信号通路激活剂LiCl则能逆转过表达UNC5A对T24细胞增殖、侵袭的抑制作用,凋亡促进作用。结论UNC5A基因过表达可能通过抑制Wnt/β-catenin信号通路的活化抑制T24细胞增殖、侵袭,并诱导细胞凋亡。

Objective To investigate the effects of UNC5A gene overexpression on the proliferation,metastasis and Wnt/β-catenin signaling pathway of bladder cancer T24 cells.Methods T24 cells were selected and transfected with UNC5A-pcDNA and pcDNA-NC,and the cells were randomly divided into three groups:control group,pcDNA-NC group and UNC5A-pcDNA group.The expression level of UNC5A mRNA was detected by RT-PCR.The rate of cell clone formation was detected by clonal formation assay.The apoptosis rate was determined by flow cytometry.Transwell detected the number of invaded cells.The expression levels of UNC5A,Wnt1 andβ-catenin were detected by Western blotting.Wnt/β-catenin signaling pathway activator LiCl was added to detect cell proliferation,apoptosis and invasion in each group.Results Compared with the control group,the expression levels of UNC5A mRNA and protein in UNC5A-pcDNA group were significantly increased(P<0.05),the rate of clone formation was decreased(P<0.05),the rate of apoptosis was increased(P<0.05),and the number of invaded cells was decreased(P<0.05).The expression levels of Wnt1 andβ-catenin protein were significantly decreased(P<0.05).The addition of LiCl,an activator of Wnt/β-catenin signaling pathway,could reverse the inhibitory effect of UNC5A overexpression on the proliferation and invasion of T24 cells and promote apoptosis.Conclusion The overexpression of UNC5A gene may inhibit the proliferation and invasion of T24 cells and induce apoptosis by inhibiting the activation of Wnt/β-catenin signaling pathway.