外源性p16基因对人原发性肝癌细胞生物学行为的影响及其机制  

The Effect and Mechanism of Exogenous p16 Expression on Biological Behavior of Human Hepatocellular Carcinoma

作　　者：罗春华[1] 朱建思[1] 苏影[1] 周建国[1] 董琳[1] 

机构地区：[1]南华大学肿瘤研究所,湖南衡阳421001

出　　处：《湖南师范大学学报（医学版）》2005年第1期24-29,共6页Journal of Hunan Normal University(Medical Sciences)

摘　　要：目的:探讨导入外源性p16cDNA真核表达载体对人原发性肝癌细胞系SMMC- 772 1生物学行为的影响及其分子机制。方法:构建外源性p16基因真核表达载体并转染SMMC- 772 1细胞,经G4 18抗性筛选和扩增,得到稳定的表达株,并经RT -PCR及免疫细胞化学鉴定。通过生长曲线,流式细胞术,免疫细胞化学及Western -Blot等实验方法,对转基因后肿瘤细胞生物学行为及细胞周期调控因子CDK4 ,CyclinD1及pRb进行观察。结果:转染外源性p16基因的SMMC- 772 1细胞有外源性p16基因的整合及表达,细胞生长速度明显减慢,倍增时间明显延长,G1期细胞明显多于转基因前(P <0 .0 5 ) ;免疫细胞化学显示外源性p16表达可下调CDK4及cylinD1的表达(P <0 .0 5 ) ,Western -Blot提示转染外源性p16基因后细胞中磷酸化pRb表达明显降低。结论:外源性p16基因导入人肝癌细胞株SMMC -772 1中可稳定表达并使细胞停滞在G1期,抑制细胞生长,其机制可能为下调CDK4、cyclinD1的表达和抑制pRb磷酸化有关。Objective To investigate the effect of exogenous p16 ex pression on b iological behavior of human hepatocellular carcinoma and to explore the mechanis m of the growth suppression of p16 in hepatocarcinoma cell line. Methods pcDNA3.1 (+)-p16 eukaryotic expression plasmid was constructed a nd transfected into SMM C-7721 cell which retains down- regulated p16 gene expression. Integration and e xpression of exogenous p16 gene were confirmed by RT-PCR and immunocytochemistr y . The curve of cell growth ra...

关 键 词：肝肿瘤 p16基因治疗 G1期阻滞 CYCLIND1 CDK4 PRB 

分 类 号：R735.7[医药卫生—肿瘤]