p16真核表达载体的构建及其对肝癌细胞的作用  

Construction of Recombinant of pcDNA 3.0/pl6 and its inhibitory role in the proliferations of hepatoma cell line

作　　者：朱争艳[1] 杜智[1] 孙铭[1] 高英堂[1] 齐之丽[1] 

机构地区：[1]天津第三中心医院,天津300170

出　　处：《临床肝胆病杂志》2003年第1期52-53,共2页Journal of Clinical Hepatology

摘　　要：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。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.

关 键 词：肝癌 CBRH-7919细胞 P16基因 基因表达 生长抑制作用 作用机制 基因治疗 

分 类 号：R735.7[医药卫生—肿瘤]