Nek2对肝癌细胞凋亡的影响及其分子机制的研究  

Effect of Nek2 on the cell apoptosis in hepatocellular carcinoma and its mechanism

作　　者：赖晓波[1] 聂玉强[1] 黄红丽[1] 李鹰飞[1] LAI Xiaobo;NIE Yuqiang;HUANG Hongli;LI Yingfei(Guangzhou Digestive Disease Center,Guangzhou First People＇ s Hospital,Guangzhou Medical University,Guangzhou 510180,China)

机构地区：[1]广州医科大学附属广州市第一人民医院消化疾病中心,广东广州510180

出　　处：《胃肠病学和肝病学杂志》2018年第6期613-616,共4页Chinese Journal of Gastroenterology and Hepatology

基　　金：广东省医学科学技术研究基金项目(A2017333)

摘　　要：目的研究Nek2(nima-related kinase2)在肝癌组织和肝癌细胞系中的表达,并通过沉默肝癌细胞中的Nek2基因,研究Nek2对肝癌细胞凋亡的影响,并初步探索其机制。方法使用Western blotting对27对肝癌及非癌组织和正常肝细胞株HL-77026及人肝癌细胞株HepG2、PLC/PRF/5、Hep3B、BEL-7402、SMMC-7721、QGY-7701中Nek2的蛋白表达情况进行分析。筛选HepG2细胞系进行下一步实验。沉默肝癌细胞HepG2中的Nek2因子,通过流式细胞仪(FACS)分析,采用Western blotting技术检测转染前后肝癌细胞中P53、Bcl-2、Bad、Caspase-3蛋白表达的变化。结果 Nek2在肝癌组织和6个肝癌细胞株中均表达升高。沉默肝癌细胞系HepG2中的Nek2后可使细胞的凋亡增强,P53、Caspase3和Bad的蛋白表达升高,抗凋亡因子Bcl-2的蛋白表达下降。结论抑制Nek2表达可以明显促进肝癌HepG2细胞的凋亡,并通过影响凋亡信号传导通路中的重要因子来促进细胞的凋亡。Objective To investigate the expression and function of nima-related kinase2（ Nek2） in hepatocellular carcinoma tissues and cell lines,and to study the apoptosis and mechanism of Nek2 in hepatocellular carcinoma. Methods The expression of Nek2 was detected by Western blotting in 27 hepatocellular carcinoma and 6 human hepatocellular carcinoma cell lines（ HepG2,PLC/PRF/5,Hep3 B,BEL-7402,SMMC-7721 and QGY-7701 cells）,to select the HepG2 cell lines for further experiments. HepG2 cells with Nek2-siRNA inhibitor or the negative control（ NC） were transfected,and the biological influence of Nek2-siRNA on HepG2 cells was analyzed in detection of apoptosis by flow cytometry and transfected by Nek2-siRNA for 48 hours,the expressions of apoptosis related factors P53,Caspase3,Bad and Bcl-2 were examined by Western blotting. Results Nek2 was upregulated in hepatocellular carcinoma tissues and cell lines.Knock down Nek2 lead to promotion of apoptosis. After knock down Nek2 in HepG2 cell,the expressions of apoptosis related factors P53,Caspase3 and Bad were increased,Bcl-2 was reduced. Conclusion Inhibiting expression of Nek2 can significantly influence the factors of cell apoptosis signal pathways by increasing P53,Caspase3,Bad,and reducing Bcl-2.

关 键 词：NEK2 肝癌 凋亡 

分 类 号：R735.7[医药卫生—肿瘤]