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目的:探讨抑癌基因p16对肝癌细胞生长的抑制作用及其机制。方法:将p16 cDNA亚克隆至pcDNA3.1真核表达载体上,并经脂质体介导转染至人肝癌细胞株SMMC-7721。用MTT法和Western blot分析转染细胞的生长情况。结果:成功构建重组表达质粒pcDNA3.1-p16,转染pcDNA3.1-p16的SMMC-7721细胞生长速度受到明显抑制;转染后有外源p16蛋白的表达,且伴随Bax上调,Bcl-2和cIAP2的下调。结论:重组pcDNA3.1-p16质粒能在人肝癌细胞SMMC-7721内表达,且能抑制SMMC-7721的生长,其机理与诱导肿瘤细胞凋亡相关。
Objective: To investigate the inhibitory effect of tumor suppressor gene p16 on the growth of hepatocellular carcinoma cells and its mechanism. Methods: The p16 cDNA was subcloned into pcDNA3.1 eukaryotic expression vector and transfected into human hepatocellular carcinoma cell line SMMC-7721 by liposome. MTT assay and Western blot analysis of transfected cells growth. Results: The recombinant plasmid pcDNA3.1-p16 was successfully constructed and the growth rate of SMMC-7721 cells transfected with pcDNA3.1-p16 was significantly inhibited. After transfection, the expression of exogenous p16 protein was up-regulated. The expression of Bcl-2 And down-regulation of cIAP2. Conclusion: The recombinant plasmid pcDNA3.1-p16 can express in human hepatocellular carcinoma cell line SMMC-7721 and inhibit the growth of SMMC-7721. Its mechanism is related to the induction of tumor cell apoptosis.