稳定表达外源性p16基因肝癌SMMC-7721细胞株的建立及鉴定  

Establishment and Identification of SMMC-7721 Cell Line With Stable Expression of Exogenous p16 Gene

作　　者：罗春华[1] 朱建思[1] 

机构地区：[1]南华大学肿瘤研究所,湖南衡阳421001

出　　处：《南华大学学报（医学版）》2005年第2期184-187,共4页Journal of Nanhua University(Medical Edition)

基　　金：湖南省卫生厅课题基金 (4 -0 2 -WS -YO2 0 68)

摘　　要：目的　构建稳定表达外源性p1 6基因的肝癌SMMC - 772 1细胞株。方法　利用脂质体介导的基因转染方法,借助真核质粒表达载体pcDNA3.1 ( + ) ,将外源性p1 6基因转染入此基因表达下调的人肝癌细胞株SMMC - 772 1细胞中,经G41 8筛选,建立稳定表达的细胞株,用逆转录聚合酶链反应(RT -PCR)及免疫组织化学法鉴定p1 6基因的表达,同时对细胞株分泌蛋白进行活性检测。结果　转染p1 6基因的SMMC - 772 1细胞中可以检测到p1 6mRNA及蛋白的表达。结论　建立稳定表达P1 6抑癌蛋白的SMMC - 772 1细胞株有助于研究p1 6抑癌基因在肝癌发生中的作用。Objective To establish SMMC-7721 cell line with stably expressing tumor suppressor gene p16. Methods The exogenous tumor suppressor gene p16 was transfected into SMMC-7721 cells with lipofectamine TM 2000 by eukaryotic expressive vector(pcDNA3.1 (+) ) . We got clone cells after selected by G418. All the clone cells were detected by RT- PCR and immunocytochemistry staining on mRNA and protein levels. Meanwhile,P16 protein activity were measured by MTT. Results p16 mRNA and protein were expressed in those transfected with p16 gene cells. Conclusion Exogenous p16 gene may stably expressed in human hepatocellular carcinoma cell line SMMC-7721. The establishment of the cell line can help to study the functions of tumor suppression gene p16 in hepatocellular carcinoma cells.

关 键 词：P16抑癌基因 基因转染 肝肿瘤 

分 类 号：R735.7[医药卫生—肿瘤]