RNAi对肝癌细胞RhoA基因表达和侵袭转移影响的研究

Effects of RhoA gene silencing on the potentiality of hepatocellular carcinoma cell metastasis

目的:探讨RNA干扰(RNAi)技术沉默RhoA基因表达对肝癌细胞HHCC生物学特性的影响。方法:合成RhoA特异性小干扰RNA(siRNA),脂质体介导转染肝癌细胞株HHCC。检测RhoA mRNA和蛋白表达水平和RhoA活性的变化,激光共聚焦显微镜观察细胞骨架变化,Boyden小室测定细胞侵袭性变化。结果:RhoA siRNA能有效抑制HHCC细胞RhoA mRNA和蛋白的表达,致使RhoA活性显著下降(P<0.01),细胞侵袭能力明显降低(P<0.01),侵袭抑制率为26.2%。结论:RhoA siRNA能够显著下调HHCC细胞RhoA的表达和侵袭能力,表明RhoA可影响肿瘤细胞的转移和侵袭能力,抑制其功能可能为肝癌治疗提供新的靶点。

OBJECTIVE：To investigate effect of RhoA gene silencing on the biological behaviors of hepatocellular carcinoma HHCC cells.METHODS：HHCC cell lines transfected with RhoA siRNA were established and gene expression was assessed by RT-PCR and Western blotting analysis.The changes of cytoskeleton were detected by laser scanning confocal microscopy,and the abilities of cell invasion and metastasis was evaluated using Boyden＇s chamber.RESULTS：RhoA siRNA effectively inhibited the expression and activity of RhoA（P0.01）.The ability of HHCC cells invasive and metastasis was depressed（P0.01）.The inhibition rate of invasion was 26.2%.CONCLUSION：RhoA siRNA can specially down-regulate RhoA expression and lead to the inhibition of malignant phenotype of HHCC cells,which further confirm that RhoA are relevant to invasion and metastatic of tumor cells and likely to be a molecular target of blocking invasion.