miR-96靶向调控NF-κB/miR-155/FOXO3a信号通路对肝癌HepG2细胞增殖和凋亡的影响  

EFFECTS OF miR-96 TARGETING REGULATION OF NF-ΚB/MIR-155/FOXO3A SIGNALING PATHWAY ON PROLIFERATION AND APOPTOSIS OF HEPATOCELLULAR CARCINOMA HEPG2 CELLS

作　　者：梅浩砚[1] 牛惠惠[1] Mei Haoyan;Niu Huihui(Department of Laboratory,The First People's Hospital of Jingmen,Jingmen 448000,China)

机构地区：[1]湖北省荆门市第一人民医院检验科

出　　处：《中国煤炭工业医学杂志》2019年第5期513-517,共5页Chinese Journal of Coal Industry Medicine

基　　金：湖北省自然科学基金项目(编号:2016CFB575)

摘　　要：目的研究miR-96靶向调控NF-κB/miR-155/FOXO3a信号通路对肝癌HepG2细胞增殖和凋亡的影响。方法经慢病毒转染沉默肝癌HepG2细胞miR-96蛋白表达,通过WST-8实验测定细胞的生长曲线,流式细胞术检测基因沉默后HeGp2的凋亡情况,Western blot分析沉默miR-96表达对肝癌细胞FOXO3a、miR-155、IKB、NF-kB、Ki67、PCNA、VEGF、Bcl-2、Bad、cleaved-caspase-3蛋白表达的影响。结果慢病毒转染后可获得遗传稳定的HepG2-miR-96-shRNA细胞系,WST-8细胞生长结果显示:与对照组比较,HepG2-miR-96-shRNA细胞的增殖能力显著降低(P <0.05);流式细胞术检测发现:HepG2-miR-96-shRNA细胞凋亡数量明显提升(P<0.05);Western blot结果提示:沉默miR-96蛋白表达后,促凋亡蛋白Bad,cleaved-caspase-3、FOXO3a及NF-κB蛋白的表达量显著增加,抑凋亡蛋白Bcl-2、miR-155、IkB及VEGF表达量明显降低。结论 miR-96通过靶向调控NF-κB/miR-155/FOXO3a信号通路抑制肝癌HepG2细胞凋亡,促进细胞增殖。Objective To study the effects of miR-96 targeting regulation of NF-κB/miR-155/FOXO3 a signaling pathway on the proliferation and apoptosis of hepatocellular carcinoma HepG2 cells.Methods The expression of miR-96 protein in silencing hepatoma HepG2 cells was detected by lentivirus transfection.The growth curve of cells was determined by WST-8 assay.The apoptosis of HeGp2 after gene silencing was detected by flow cytometry.Western blot analysis was used to analyze the effects of silencing miR-96 expression on the protein expression levels of FOXO3 a,miR-155,IKB,NF-κB,Ki67,PCNA,VEGF,Bcl-2,Bad and cleaved-caspase-3.Results Western blot After lentivirus transfection,HepG2-miR-96-shRNA cell lines with genetic stability were obtained.The growth of WST-8 cells showed that compared with the control group,the proliferation of HepG2-miR-96-shRNA cells significantly reduced(P<0.05).Flow cytometry showed that the apoptotic number of HepG2-miR-96-shRNA cells significantly increased(P<0.05).The results of Western blot showed that after silencing the expression of miR-96 protein,the expression of pro-apoptotic proteins Bad,cleaved-caspase-3,FOXO3 a and NF-κB increased significantly,while the expression of anti-apoptotic proteins Bcl-2,miR-155,IkB and VEGF decreased significantly.Conclusion miR-96 inhibits apoptosis and promotes proliferation of HepG2 cells by the targeting regulation of NF-κB/miR-155/FOXO3 a signaling pathway.

关 键 词：miR-96 靶向调控 NF-κB信号通路 FOXO3A 细胞凋亡 

分 类 号：R735.7[医药卫生—肿瘤]