摘要
目的探讨miR-106b及核心亚基基因增强子的人同源基因2(EZH2)对HepG2细胞生长的影响及其作用机制。方法 qRT-PCR法检测不同肝癌细胞miR-106b的表达;用miR-106b抑制物(inhibitor)及模拟物(mimic)和siEZH2转染肝癌细胞,CCK8法检测细胞增殖,细胞凋亡用Annexin V-FITC/PI双染法和WB法检测凋亡蛋白-3(Caspase-3),生物信息学分析并用荧光素酶法证实miR-106b是否靶向结合EZH2。结果与永生化的肝细胞LO2相比,miR-106b在肝癌细胞中表达量增加;miR-106b mimic促进HepG2细胞增殖和抑制凋亡;miR-106b inhibitor能导致Caspase-3全长表达量减少和其活性片段表达量增加;siEZH2抑制HepG2细胞增殖和促进其凋亡,并且能导致Caspase-3全长表达量减少及其活性片段表达量增加。荧光素酶结果表明miR-106b并非直接靶向抑制EZH2,miR-106b促进EZH2的表达。结论抑制miR-106b和EZH2的表达都能有效抑制HepG2细胞增殖和促进其凋亡。
Objective To study of effects and mechanisms of miR-106b and EZH2 on HepG2 cell proliferation. Methods The expression of miR-106b was detected by q RT-PCR in liver cancer cells. Cell Counting Kit-8(CCK-8)was used as the cell proliferation assay. Cell apoptosis was detected by Annexin V-FITC/PI double staining and WB. The relationship between miR-106b and EZH2 was assessed by bioinformatics analysis and Dual-Luciferase Reporter assay. Results The expression of miR-106b increased in liver cancer cells. miR-106b promoted proliferation of HepG2 cells and inhibited apoptosis. miR-106b inhibitor suppressed the expression of full length of Caspase-3 and promoted the expression of its cleaved fragments. siEZH2 inhibited the proliferation of HepG2 cells and promoted their apoptosis,and suppressed the expression of full length of Caspase-3 and promoted the expression of its cleaved fragment in HepG2 cells. EZH2 was not a direct target of miR-106b by Dual-Luciferase Reporter assay;on the contrary,miR-106b promoted the expression of EZH2.Conclusion Inhibition of miR-106b and EZH2 could inhibit the proliferation of HepG2 cells and promote apoptosis.
作者
桂连
张倩倩
蔡燕
邓小红
黄俊琪
GUI Lian ZHANG Qian-qian CAI Yan DENG Xiao-hong HUANG Jun-qi(Guangdong Provincial Key Laboratory of Organ Donation and Transplant Immunology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510080 Guangdong Provincial International Cooperation Base of Science and Technology (Organ Transplantation), The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510080 1nstitute of lmmunology , Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, Guangdong 510080, China)
出处
《热带医学杂志》
CAS
2017年第10期1279-1284,共6页
Journal of Tropical Medicine
基金
国家自然科学基金(31370870)
广东省自然科学基金(S2012010009050
S2013020013000)
广东省科技计划项目(2010B050700008
2011B040300022
2013A02022 9003)
广州市科技计划项目(11C32060749
2011J4100084
201604020083)
广东省器官捐献与移植免疫重点实验室建设项目(2013A061401007)
广东省器官移植国合基地建设项目(2015B050501002)
2010年中山大学高校基本科研业务费青年培育项目(10ykpy31)