β-catenin在膀胱癌上皮-间质转化中的作用及相关机制

The role and mechanism of β-catenin in epithelial-mesenchymal transition in bladder cancer

目的:探讨β-catenin在膀胱癌上皮-间质转化(epithelial-mesenchymal transition,EMT)过程中的作用以及相关分子机制。方法:利用siRNA靶向沉默膀胱癌细胞系T24细胞的β-catenin基因,并利用Real-time quantitative PCR和Western blot实验验证siRNA敲减效率。通过Transwell实验检测β-catenin敲减后T24细胞的侵袭能力,进一步使用Real-time quantitative PCR和Western blot实验检测β-catenin敲减后T24细胞的EMT相关标志物的变化。最后,通过下载TCGA数据库,并分析β-catenin与EMT相关标志物的相关性。结果:我们成功在mRNA和蛋白水平上沉默了β-catenin。通过Transwell实验发现,与对照组相比,β-catenin敲减组细胞的侵袭能力下调。与对照组相比,β-catenin敲减组的Slug蛋白、Vimentin蛋白和mRNA下调,E-cadherin蛋白和mRNA上调。通过分析TCGA数据库发现,β-catenin与Slug(r=0.183 5,P=0.000 2)和Vimentin(r=0.190 3,P=0.000 1)均呈正相关。结论:β-catenin通过调控Slug的表达从而调控膀胱癌细胞的EMT。

Objective：To investigate the role of β-catenin in the process of EMT in bladder cancer.Methods：We established β-catenin knockdown T24 cell lines using siRNA strategy and detected the knockdown efficiency ofβ-catenin using real-time quantitative PCR and Western blot.We performed Transwell experiment to evaluate the invasion of β-catenin-silencing T24 cell lines.Furthermore,we detected the expression of EMT markers by Real-time quantitative PCR and Western blot.Finally,we downloaded the public TCGA database to analyze the correlation between β-catenin and EMT markers.Results：We established the β-catenin knockdown T24 cell lines successfully.Compared with negative control（ NC）,β-catenin knockdown T24 cells showed decreased invasion.Moreover,compared with NC,the epithelial marker（ E-cadherin） increased and mesenchymal markers（ Slug and Vimentin）decreased in β-catenin knockdown T24 cells.By analyzing the TCGA database,we found that β-catenin showed positive correlation with Slug（ r = 0.183 5,P = 0.000 2） and Vimentin（ r = 0.190 3,P = 0.000 1）.Conclusion：β-catenin could modulate EMT in bladder cancer by targeting Slug.