miR-363靶向调控E2F3表达影响HepG2细胞增殖和凋亡研究  

Influence of miR-363 on the proliferation and apoptosis of HepG2 cells by targeting E2F3 in vitro

作　　者：姜哲 舒敏[1] 王媛媛[1] 朴莲淑[1] Jiang Zhe;Shu Min;Wang Yuanyuan(Department of Gastroenterology,Zhongshan Hospital,Dalian University,Dalian 116000,Liaoning Province,China)

机构地区：[1]大连大学附属中山医院消化内科,辽宁省大连市116000

出　　处：《实用肝脏病杂志》2018年第6期825-828,共4页Journal of Practical Hepatology

基　　金：国家自然科学基金资助项目(编号:81301979)

摘　　要：目的探讨微小RNA-363(miR-363)靶向调控E2F转录因子3(E2F3)的表达对HepG2细胞增殖和凋亡的影响。方法体外培养HepG2细胞,采用Lipofectamine法将miR-363抑制剂或其阴性对照转染到HepG2细胞,继续培养48 h,收获细胞,采用四噻唑蓝(MTT)法测定细胞增殖率,使用流式细胞术法检测细胞凋亡率,采用实时荧光RT-PCR法检测HepG2细胞miR-363 mRNA水平,采用Western Blot法检测HepG2细胞E2F3、BAX和Caspase-3蛋白表达水平。结果对照组HepG2细胞增殖率为(96.4±9.7)%,显著高于抑制剂处理组[(72.3±6.5)%,P<0.05],凋亡率为(8.2±1.4)%,显著低于抑制剂处理组[(9.7±0.8)%,P<0.05];对照组HepG2细胞miR-363 mRNA相对水平为(1.0±0.1),显著高于抑制剂处理组[(0.6±0.2),P<0.05],E2F3蛋白表达量为(1.0±0.1),显著高于抑制剂处理组[(0.6±0.1),P <0.05],而对照组HepG2细胞Bax和Caspase-3蛋白表达量分别为(0.4±0.0)和(0.5±0.1),均显著低于抑制剂处理组[(0.6±0.1)和(0.7±0.0),P均<0.05]。结论 miR-363可靶向调控E2F3的表达,抑制HepG2细胞增殖,诱导其凋亡。本研究结果为肝癌靶向治疗提供了一定的理论依据。Objective To investigate the influence of miR-363 on the proliferation and apoptosis of HepG2 cells by targeting E2F3 in vitro.Methods The HepG2 cells was cultured and transfected with miR-373 inhibitors or negative control(NC)by Lipofectamine liposome method.At 48 h,MTT method and flow cytometry were used to detect the cell proliferation and apoptosis respectively.The miR-363 mRNA in HepG2 cells was detected by RT-PCR and the E2F3,BAX and caspase-3 proteins in HepG2 cells were detected by Western blot.Results The proliferation rate of HepG2 cells in the control group was(96.4±9.7)%,significantly higher than[(72.3±6.5)%,P<0.05]in the inhibitor-intervened group,while the apoptosis rate was(8.2±1.4)%,significantly lower than[(9.7±0.8)%,P<0.05]in the inhibitor-intervened group;the relative miR-363 mRNA level was(1.0±0.1),significantly higher than[(0.6±0.2),P<0.05]in the inhibitor-intervened group;the expression of E2F3 protein was(1.0±0.1),significantly higher than[(0.6±0.1),P<0.05],while the expression of Bax and caspase-3 protein were(0.4±0.0)and(0.5依0.1),significantly lower than[(0.6±0.1)and(0.7±0.0),respectively,P<0.05]in the inhibitor-intervened group.Conclusion The miR-363 can inhibit the proliferation,and induce the apoptosis of HepG2 cells by targeting E2F3,which provides a theoretical basis for targeted therapy of liver cancer.

关 键 词：HEPG2细胞 微小RNA-363 E2F转录因子 细胞增殖 细胞凋亡 

分 类 号：R735.7[医药卫生—肿瘤]