p16真核表达载体的构建及其对肝癌细胞的作用

研究p16基因对肝癌细胞的作用;构建pcDNA3.0/p16真核表达质粒转导到大鼠肝癌细胞CBRH-7919中,对其p16基因的表达、细胞的生长抑制及机制进行分析;转染细胞p16蛋白免疫组化阳性,MTY法结果显示,50×103/cm2细胞经培养24h～96h后,每组细胞数均有不同程度的增加,但经pcDNA3.0/p16转染的CBRH-7919细胞数比对照明显减少。与空白对照比,细胞在24h,48h,72h和96h的生长抑制率分别为29％,29％,40％和52％,流式细胞仪检测细胞周期显示经pcDNA3.0/p16转染的CBRH-7919细胞有显著的细胞凋亡现象和G0/G1期阻滞。pcD-NA3.0/p16真核表达质粒转导到大鼠肝癌细胞CBRH-7919中能够抑制细胞的增殖活性,p16基因可能通过诱导肿瘤细胞凋亡及G1期阻滞在肿瘤的基因治疗方面发挥作用。

Construction of Recombinant of pcDNA 3.0/p16 and its inhibitory role in the proliferations of hepatoma cell line

To explore the suppressive function of wild type p16 gene in the growth of hepatoma cell. p16 cDNA was subcloned into pcDNA 3.0. And pcDNA 3.0/pl6 eukaryotic expression plasmid was constructed. Its inhibitory role in the proliferation of rat hepatoma cell line CBRH - 7919 was investigated. MTT method and flow cytometry were used to determine the cell viability and cell cycle of the transfected CBRH - 7919 cells. pcDNA 3.0/pl6 eukaryotic expression plasmid was obtained and the transfected cells could express p16 protein. The viability of the transfected CBRH - 7919 cells was decreased and the percentage of the cell in G0/G1 phase was increased. pcDNA 3.0/pl6 plasmid could be expressed in CBRH - 7919 cells and inhibit the proliferation of the CBRH - 7919. P16 gene may play an important role on gene therapy of tumors by the induction of tumor cells apotosis and the blockage of cellular G1 stage.