小干扰RNA沉默CIP2A表达阻滞HepG2细胞周期、促进细胞衰老的作用及机制探讨  

Silencing of CIP2A expression by siRNA blocks HepG2 cell cycle and promotes senescence and its molecular mechanism

作　　者：杨雪[1] 张靖 韩少山[1] 陶杰[1] 孙昊[1] 刘青光[1] Yang Xue;Zhang Jing;Han Shaoshan;Tao Jie;Sun Hao;Liu Qingguang(Department of Hepatobiliary Surgery ,the First Affiliated Hospital of Xi'an Jiaotong University ,Shaanxi Xi'an 710061 ,China;Department of Gynecologic Oncology , Shaanxi Provincial Cancer Hospital , Shaanxi Xi'an 710061 , China)

机构地区：[1]西安交通大学第一附属医院肝胆外科,陕西西安710061 [2]陕西省肿瘤医院妇瘤病院,陕西西安710061

出　　处：《现代肿瘤医学》2019年第12期2051-2054,共4页Journal of Modern Oncology

基　　金：西安交通大学第一附属医院科研基金青年创新项目(编号:2017QN-19)

摘　　要：目的:观察抑制CIP2A表达对HepG2细胞周期及衰老的影响,并探讨初步机制。方法:运用前期成功构建的重组腺相关病毒载体rAAV2-EGFP-CIP2A shRNA转染肝癌细胞系HepG2,PI单染测定细胞周期,衰老相关β-半乳糖苷酶染色检测阳性细胞率,Western blot检测细胞周期相关蛋白的表达。结果:病毒感染组与空病毒感染组细胞相比,抑制CIP2A表达后HepG2细胞出现G_1期阻滞(69.8%vs 60.4%,P=0.025),细胞内β-半乳糖苷酶染色阳性比例升高(22.1%vs 9.6%,P=0.022),细胞周期相关蛋白FoxM1、CDK2、CDK4、cyclin D1及MCM7蛋白表达量降低,而细胞周期负性调控分子p21蛋白表达量增加。结论:抑制CIP2A表达导致HepG2细胞出现细胞周期停滞,细胞衰老增加,其机制可能通过调控细胞周期相关蛋白表达来实现的。Objective:To investigate the effect of inhibiting CIP2 A expression on cell cycle and senescence of HepG2 in hepatocellular carcinoma cells,and explore the preliminary mechanism.Methods:The recombinant adeno-associated viral vector rAAV2-EGFP-CIP2 A shRNA was successfully transfected into HepG2 in HCC cell lines by using previous experiments.PI staining was used to determine the cell cycle.Positive cell rate was detected by beta-galactosidase staining,and the expression of cyclin-related proteins was detected by Western blot.Results:Compared with the control group,HepG2 cells in the viral infection group showed G1 phase arrest after inhibiting CIP2 A expression(69.8% vs 60.4%,P=0.025),the positive proportion of intracellular beta-galactosidase staining increased(22.1% vs 9.6%,P=0.022),and the expression levels of cyclin-related proteins FoxM1,CDK2,CDK4,cyclin D1 and MCM7 decreased,while the expression levels of p21 increased.Conclusion:Inhibition of CIP2 A expression leads to cell cycle arrest and increased cell senescence in HepG2 cells,which may be achieved by regulating the expression of cyclin-related proteins.

关 键 词：CIP2A 肝癌 细胞周期 细胞衰老 

分 类 号：R735.7[医药卫生—肿瘤]