基于肝癌CDK2基因敲减对CyclinE表达及树舌多糖GF对其影响的研究  

Study on the Effect of CDK2 Knockdown Decreases on the Expression of CyclinE and GAPS.GF in Hepatocellular Carcinoma

作　　者：于英君[1] 韩松洋[1] 姜颖[1] 张梅[1] 杨丹[1] 葛圆圆[1] 于水澜[1] 

机构地区：[1]黑龙江中医药大学基础医学院,黑龙江哈尔滨150040

出　　处：《现代生物医学进展》2016年第23期4434-4437,4461,共5页Progress in Modern Biomedicine

基　　金：黑龙江省中医管理局项目(ZHY06-Z02);黑龙江中医药大学科研基金项目(X201004);黑龙江中医药大学科研基金项目(201409);黑龙江省博士后基金项目(LBH-Z15211)

摘　　要：目的:探究树舌多糖GF(GAPS.GF)对rAAV-shRNA-CDK2抑制肝癌细胞Cyclin E基因表达的辅助作用。方法:将细胞培养后的人肝癌HepG2以皮下注射的方式接种于裸鼠前肢腋下,接种后的裸鼠随机分为5组:NC(非相关序列)对照组、肿瘤组、rAAV-shRNA-CDK2组、树舌多糖GF组以及树舌多糖GF+rAAV-shRNA-CDK2组,各实验组均采取尾静脉注射定量给药。采用实时定量PCR和Western blot技术研究肝癌细胞Cyclin E基因m RNA和蛋白水平的表达情况,同时观察GASP.GF对其作用的影响。结果:树舌多糖GF+rAAV-shRNA-CDK2联合应用组对肝癌HepG2细胞增殖的抑瘤率为75.6%,对Cyclin E基因mRNA表达抑制率为69%,对Cyclin E基因蛋白表达抑制率为67.5%,比rAAV-shRNA-CDK2组分别提高了3.42%、1%和2.7%。结论:树舌多糖GF与rAAV-shRNA-CDK2联合应用可以显著提高肝癌的治疗效果,说明树舌多糖GF可以辅助r AAV-shRNACDK2对肝癌细胞Cyclin E基因的表达的抑制作用。Objective： To study the auxiliary function of GAPS.GF on inhibition the Cyclin E gene expression in hepatocellular carcinoma cell by r AAV-sh RNA-CDK2. Methods： Human hepatocellular carcinoma HepG2 cells were cultured and inoculated into nude mice by subcutaneous injection. The nude mice were randomly divided into 5 groups： a NC contrast group, a tumor group, a r AAV-sh RNA-CDK2 group, a GAPS.GF group and a GAPS.GF＋ r AAV-sh RNA-CDK2 group, all the experimental groups were administered by caudal vein. Using Real-time PCR and Western blot to study the expression of m RNA and protein levels of Cyclin E gene in hepatocellular carcinoma cells and then observing the effect of GAPS.GF on its function. Results： The inhibition rate of proliferation of hepatocellular carcinoma HepG2 cells was 75.6 % in the combination group of GAPS.GF＋r AAV-sh RNA-CDK2 group, the inhibition rate of Cyclin E gene m RNA expression was 69 %, the inhibition rate of Cyclin E gene protein expression was 67.5 %, than the r AAV-sh RNA-CDK2 group were increased by 3.42 %, 1 %, 2.7 %. Conclusions： The combined application of GAPS.GF and r AAV-sh RNA-CDK2 can significantly improved the treatment effect of liver cancer, which shows that the GAPS.GF can help r AAV-sh RNA-CDK2 to inhibit the expression of Cyclin E gene in hepatocellular carcinoma cells.

关 键 词：肝癌HEPG2细胞 rAAV-shRNA-CDK2 树舌多糖GF(GAPS.GF) Cyclin E 

分 类 号：R735.7[医药卫生—肿瘤]