miR-363靶向调控E2F3表达影响HepG2细胞增殖和凋亡研究*

doi:10.3969/j.issn.1672-5069.2018.06.001

实用肝脏病杂志 ›› 2018, Vol. 21 ›› Issue (6): 825-828.doi: 10.3969/j.issn.1672-5069.2018.06.001

• 实验性肝炎 • 下一篇

miR-363靶向调控E2F3表达影响HepG2细胞增殖和凋亡研究^*

姜哲, 舒敏, 王媛媛, 朴莲淑

116000 辽宁省大连市大连大学附属中山医院消化内科

收稿日期:2018-03-26 出版日期:2018-11-10 发布日期:2018-12-25
通讯作者: 朴莲淑,E-mail:lianshupiao0627@hotmail.com
作者简介:姜哲,男,36岁,大学本科,主治医师。研究方向：主要从事消化内科疾病防治研究。E-mail：113239230@qq.com
基金资助:
*国家自然科学基金资助项目（编号：81301979）

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

Jiang Zhe, Shu Min, Wang Yuanyuan, et al.

Department of Gastroenterology,Zhongshan Hospital,Dalian University, Dalian 116000,Liaoning Province,China

Received:2018-03-26 Online:2018-11-10 Published:2018-12-25

摘要/Abstract

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。

关键词: HepG2细胞, 微小RNA-363, E2F转录因子, 细胞增殖, 细胞凋亡

Abstract: 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±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.

Key words: HepG2 cells, miR-363, E2F transcription factor, Proliferation, Apoptosis

姜哲, 舒敏, 王媛媛, 朴莲淑. miR-363靶向调控E2F3表达影响HepG2细胞增殖和凋亡研究^*[J]. 实用肝脏病杂志, 2018, 21(6): 825-828.

Jiang Zhe, Shu Min, Wang Yuanyuan, et al.. Influence of miR-363 on the proliferation and apoptosis of HepG2 cells by targeting E2F3 in vitro[J]. JOURNAL OF PRACTICAL HEPATOLOGY, 2018, 21(6): 825-828.

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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

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