Nodal靶向RNA干扰对人肝癌细胞的影响

Effect of nodal targeting RNA interference on human hepatocellular carcinoma cells

目的:观察RNA干扰技术对人肝癌细胞Nodal基因表达的干扰效应,探讨靶向Nodal基因在肝癌基因治疗中的可行性。方法:采用一段含针对Nodal特异shRNA序列的质粒载体,转染人肝癌细胞株SMMC-7721,观察其转染效率后,分别以实时荧光定量PCR和Western blot检测Nodal基因和蛋白的表达水平,并观察Nodal基因干扰对SMMC-7721细胞体外增殖的影响。结果:表达shRNA的质粒载体对SMMC-7721细胞有较高的转染效率,转染后可明显抑制SMMC-7721细胞Nodal基因和蛋白的表达,且对其体外增殖有明显抑制作用。结论:运用RNA干扰技术,可以有效地抑制肝癌细胞Nodal基因和蛋白的表达,并抑制细胞增殖,Nodal基因可望成为肝癌治疗的新靶点。

Objective： To observe the inhibitory effect of RNA interference （RNAi） on the expression of nodal gene in human hepatocellular carcinoma （HCC） cells, and explore the feasibility of nodal gene targeting in HCC gene therapy. Methods： SMMC-7721 cells were transfected with the plasmid vector containing a short hairpin RNA （shRNA） sequence targeting the nodal gene. After assessing the transfection efficiency, the nodal gene and protein expression was detected by real-time quantitive PCR and Western blot, respectively. Furthermore, the effect of nodal gene interference on the proliferation of SMMC-7721 cells in vitro was studied. Results： The plasmid containing shRNA had excellent transfection efficiency in SMMC-7721 ceils. After transfection, the expression of nodal mRNA and protein of SMMC-7721 cells was significantly inhibited, and their proliferation in vitro was also significantly suppressed.Conclusion： The nodal gene and protein expression as well as cell proliferation of HCC cells can be effectively inhibited by RNAi technique, so nodal gene may be a potential therapeutic target for HCC.