LIM激酶1对肝癌细胞增殖与转移的调控作用

Regulatory effects of LIM kinase 1 on the proliferation and metastasis of hepatocellular carcinoma cells

目的研究LIM激酶1(LIMK1)在肝癌组织及细胞中的表达情况,以及LIMK1对肝癌细胞增殖与转移的调控作用。方法通过在线数据库starBase v3.0和GEPIA分析LIMK1在肝癌组织和肝脏正常组织中的表达情况,并进行相关生存分析;蛋白质印迹(Western blot)技术分析LIMK1在肝癌细胞株中的表达情况;用小干扰RNA(siRNA)技术下调LIMK1的表达,瞬时转染肝癌细胞Hep3B和Huh7,通过四甲基偶氮唑盐实验及克隆形成实验观察其对肝癌细胞增殖的调控作用;Transwell实验检测LIMK1的表达下调后肝癌细胞转移能力的变化;下调LIMK1的表达后,Western blot技术检测上皮-间质转化进程中相关指标的改变。对数据进行单因素方差分析。结果LIMK1在肝癌组织中的表达水平明显高于正常肝组织,而且与预后相关(P值均<0.01);进一步研究表明,与永生化的肝细胞L02相比,LIMK1在肝癌细胞株中的表达明显升高(P值均<0.05)。功能相关实验表明,在LIMK1表达下调后,肝癌细胞的增殖与转移能力明显受到抑制(P值均<0.05);同时发现LIMK1参与上皮-间质转化进程。结论LIMK1在肝癌组织及细胞中表达水平明显升高,可调控肝癌细胞的增殖与转移并且参与上皮-间质转化进程。

Objective To study LIM kinase 1(LIMK1)expressional condition,and its regulatory effects on the proliferation and metastasis of hepatocellular carcinoma cells and tissues.Methods The online database starBase v3.0 and GEPIA were used to analyze the LIMK1 expression in hepatocellular carcinoma cells and normal liver tissues,and then the relevant survival analysis was performed.LIMK1 expression in hepatocellular carcinoma cell line was analyzed by Western blot.Hep3B and Huh7 cells were transiently transfected after LIMK1 protein expression was down-regulated by small interfering RNA(siRNA).LIMK1 effects on the proliferation of Hep3B and Huh7 cells were observed by MTT assay and colony formation assay.Transwell assay was used to detect the change in metastatic ability of hepatocellular carcinoma cell after the down-regulation of LIMK1 expression.Western blot was used to detect the changes of related indexes in the process of epithelial mesenchymal transition after the down-regulation of LIMK1 expression.Data were analyzed by one-way ANOVA.Results The expression level of LIMK1 in liver cancer tissues was significantly higher than that of normal liver tissues,and was related with prognosis(P<0.01).Furthermore,LIMK1 expression in HCC cell lines was significantly higher than that of immortalized liver L02 cells(P<0.05).Functional correlated experiment showed that the proliferation and metastatic ability of liver cancer cells were significantly inhibited after LIMK1 expression down-regulation(P<0.05).Simultaneously,LIMK1 was also involved in the process of epithelial-mesenchymal transition.Conclusion LIMK1 was overexpressed in HCC tissues and cells,and may regulate the proliferation and metastasis of HCC cells and participate in epithelial-mesenchymal transition process.