IGF-Ⅱ对人肝癌细胞HepG2中抑癌基因p16和p53表达的影响  

The influence of IGF-Ⅱ on expressions of antioncogenes p16 and p53 in HepG2 cells

作　　者：姬媛媛[1] 王志东[2] 陈海燕[1] 王宝太[2] 杨正安[2] 惠博[2] 史敏[2] 卓飞[2] 

机构地区：[1]西安交通大学第二附属医院科研中心实验室 [2]西安交通大学第二附属医院干四病区老年消化外科,陕西西安710004

出　　处：《现代肿瘤医学》2017年第12期1853-1858,共6页Journal of Modern Oncology

基　　金：国家自然科学基金资助项目(编号:81201925);西安交通大学第二附属医院人才培养专项科研基金资助项目[编号:RC(GG)201502]

摘　　要：目的:观察胰岛素样生长因子-Ⅱ(IGF-Ⅱ)对人肝癌细胞HepG2中生物学表型变化及抑癌基因p16和p53表达的影响。方法:培养人肝癌细胞株HepG2,IGF-Ⅱ(25、50、100 ng/ml)作用于不同时间点(24、48、72 h),观察各组细胞生物学表型变化。IGF-Ⅱ(50 ng/ml)作用于HepG2 48 h后,分别采用Realtime PCR、Western blot及细胞免疫荧光法检测人肝癌细胞中p16和p53的表达情况,并进行定量分析。结果:IGF-Ⅱ诱导HepG2细胞形态发生一定改变;IGF-Ⅱ可增加HepG2细胞存活率及细胞增殖,但差异无统计学意义(P>0.05);IGF-Ⅱ(50 ng/ml)对HepG2细胞的克隆形成有一定的促进作用,但差异无统计学意义(P>0.05)。与对照组相比,IGF-Ⅱ组HepG2细胞中p16和p53 mRNA及蛋白表达量明显降低(P<0.05)。此外,p16和p53阳性表达主要位于细胞核。对照组、IGF-Ⅱ组HepG2细胞中p16荧光表达量分别为(100.00±11.55)%、(68.33±6.01)%,p53荧光表达量分别为(100.00±17.32)%、(57.67±10.11)%。与对照组相比,IGF-Ⅱ组HepG2细胞中p16及p53荧光表达量明显减弱(P<0.05)。结论:IGF-Ⅱ可通过诱导人肝癌细胞HepG2生物学表型的改变,下调抑癌基因p16和p53的mRNA及蛋白表达,继而发挥促进肝癌进展的作用。Objective： To probe the influence of IGF-Ⅱ on changes of biological phenotype and expressions of antioncogenes p16 and p53 in HepG2 cells. Methods： HepG2 cells were cultured and stimulated with IGF-Ⅱ（ 25,50 and 100 ng/ml） for 24,48 and 72 h,changes of biological phenotype were observed. And HepG2 cells were stimulated with IGF-Ⅱ（ 50 ng/ml） for 48 h,mRNA levels and protein expressions of antioncogenes p16 and p53 were examined by Real-time PCR and Western blot. Moreover,expressions of p16 and p53 were detected by immunofluorescence. Results： HepG2 cells induced by IGF-Ⅱ showed a morphological alteration. MTT,Brdu and clone-formation assay suggested that growth and proliferation of HepG2 cells were enhanced by IGF-Ⅱ,but no significant difference was found between IGF-Ⅱ groups with control group（ P 〉0. 05）. Compared with the control,mRNA levels of p16 and p53 were significantly decreased by IGF-Ⅱ（ P〈0. 05） in HepG2 cells,and protein expressions of p16 and p53 were remarkably reduced by IGF-Ⅱ（ P〈0. 05） in HepG2 cells. Furthermore,immunofluorescent expressions of p16 and p53 could be found in the nucleus of HepG2 cells. Compared with the control,immunofluorescent expressions of p16 and p53 were distinctly declined by IGF-Ⅱ in HepG2 cells（ P〈0. 05）. Conclusion： IGF-Ⅱ might induce changes of biological phenotype and inhibit expressions of antioncogenes p16 and p53 in HepG2 cells,further promoting the development of hepatocellular carcinoma.

关 键 词：IGF-Ⅱ p16 p53 生长增殖 HEPG2细胞 

分 类 号：R735.7[医药卫生—肿瘤]