CTCF通过抑制p53信号通路促进胆管癌细胞的生长、迁移和侵袭

CTCF promotes the growth, migration and invasion ofcholangiocarcinoma cells by inhibiting the p53 signaling pathway

为了探索CCCTC结合因子(CCCTC-binding factor,CTCF)对人胆管癌细胞的生长、迁移和侵袭能力的影响及其潜在的分子作用机制,利用慢病毒感染法获得稳定过表达或敲低CTCF的胆管癌细胞系,采用蛋白质免疫印迹法(Western blotting,Wb)检测CTCF蛋白表达水平,采用Cell Counting Kit-8(CCK-8)法和克隆形成实验检测细胞生长情况,采用Transwell小室实验检测细胞迁移和侵袭能力,采用GraphPad软件(v6)的Mann-Whitney U检验分析CTCF基因在癌和癌旁组织间的mRNA差异表达.结果显示:过表达CTCF可显著促进HCCC-9810和RBE胆管癌细胞的生长、迁移和侵袭能力;敲低CTCF呈现相反的表型.通过通路富集分析发现:CTCF的表达水平在胆管癌中与p53信号通路活性显著负相关,过表达CTCF可显著下调p53及其靶基因p21的mRNA和蛋白表达水平,而敲低CTCF则显著促进p53和p21的mRNA和蛋白表达水平.综上,本研究揭示CTCF可能通过抑制p53信号通路而促进胆管癌细胞生长、迁移和侵袭而发挥促癌基因的功能.

The study aims to explore the functions and potential molecular mechanisms of CCCTC-binding factor(CTCF)in cholangiocarcinoma(CCA)cell lines.The CCA cell lines with stable overexpression or knockdown of CTCF were obtained by lentivirus infection.Western blotting(Wb)assays were used to detect the CTCF protein expression levels.The cell growth was detected by CCK-8 assays and plated cell clone formation assays.Cell migration and invasion was detected by Transwell.Mann-Whitney U test of GraphPad software (v6) was used to analyze the mRNA differential expression of CTCF gene between cancer and adjacent tissues. The results show that overexpression of CTCF significantly promoted the growth, migration and invasion of HCCC-9810 and RBE cells, whereas knockdown of CTCF exhibited the opposite effects. Through pathway enrichment analysis, it was found that the expression level of CTCF was significantly negatively correlated with the activity of p53 signaling pathway in cholangiocarcinoma.Overexpression of CTCF significantly reduced the expression of p53 and its target gene p21 , while knockdown of CTCF showed the opposite effects. In conclusion, this study revealed that CTCF function as an oncogene by inhibiting the p53 signaling pathway to promote the growth, migration and invasion of CCA cells.