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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 interference effect of RNAi technology on the expression of Nodal gene in human hepatocellular carcinoma cells and to explore the feasibility of targeting Nodal gene in hepatocellular carcinoma gene therapy. Methods: A plasmid vector containing Nodal-specific shRNA sequence was transfected into human hepatocellular carcinoma cell line SMMC-7721. The transfection efficiency was observed. The expression of Nodal gene and protein was detected by real-time fluorescence quantitative PCR and Western blot respectively. To observe the effect of Nodal gene interference on the proliferation of SMMC-7721 cells in vitro. Results: The plasmid vector expressing shRNA had a higher transfection efficiency on SMMC-7721 cells. The transfection could obviously inhibit the expression of Nodal gene and protein in SMMC-7721 cells and significantly inhibit the proliferation of SMMC-7721 cells. Conclusion: The RNA interference technique can effectively inhibit the expression of Nodal gene and protein in hepatocellular carcinoma cells and inhibit cell proliferation. Nodal gene is expected to become a new target for the treatment of liver cancer.