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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 the effect of silencing RhoA gene expression on biological characteristics of hepatocellular carcinoma HHCC by RNA interference (RNAi) technique. Methods: RhoA specific small interfering RNA (siRNA) was synthesized and lipofectamine was transfected into hepatocellular carcinoma cell line HHCC. The changes of RhoA mRNA and protein expression and RhoA activity were detected. The change of cytoskeleton was observed by laser confocal microscopy. The invasiveness of cells was determined by Boyden chamber. Results: RhoA siRNA could effectively inhibit the expression of RhoA mRNA and protein in HHCC cells, resulting in a significant decrease in RhoA activity (P <0.01) and a significant decrease in invasion ability (P <0.01). The invasion inhibition rate was 26.2%. Conclusion: RhoA siRNA can down-regulate RhoA expression and invasion ability of HHCC cells significantly, indicating that RhoA can affect the metastasis and invasion ability of tumor cells. Inhibiting its function may provide a new target for HCC treatment.