p16基因对人肝癌细胞的生长抑制作用的初步研究

The growth inhibitory effects by transfection of p16 gene on human hepatic cancer cell line*

目的探讨抑癌基因p16对肝癌细胞生长的抑制作用。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;经Western blot证实,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。

Objectiv p16 gene is a tumor suppressor gene and is inactivated in the carcinogenesis of hepatic cancer. This paper was to test ability to inhibit growth of hepatic cancer cell of p16 gene. Method p16 cDNA was subcloned to pcDNA3.1 eucaryotic expressing vector; then the recombinant pcDNA3.1-p16 plasmi plasmid was transfected into human hepatic cell line SMMC-7721 with liposome. MTT method and Western blot were employed to investigate cell growth viability. Results The recombinant plasmid pcDNA3.1-p16 was built successfully; the growth of cancer cell line SMMC-7721 was obviously inhibited after transfection; the exogenous p16 protein was expressed increasedly with the upregulation of Bax： and downregulation of Bcl-2 and cIAP2 by Western blot analysis. Conclusion The recombinant plasmid pcDNA3.1-p16 is able to express in human hepatic cell line SMMC-7721 and suppress the cancer cell growth, which is associated with apoptosis.