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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 investigate the effect of human cognate gene 2 (EZH2) with miR-106b and core subunit enhancer on HepG2 cell growth and its mechanism. Methods The expression of miR-106b in different hepatocellular carcinoma cells was detected by qRT-PCR. The hepatocellular carcinoma cells were transfected with miR-106b inhibitor and mimic and siEZH2. The cell proliferation and apoptosis were detected by Annexin V- FITC / PI double staining and WB assay Caspase-3, bioinformatics analysis and luciferase confirmed whether miR-106b targeted binding to EZH2. Results Compared with immortalized LO2 cells, miR-106b expression was increased in HepG2 cells; miR-106b mimic promoted HepG2 cell proliferation and inhibited apoptosis; miR-106b inhibitor led to the decrease of full-length Caspase-3 expression and The expression of the active fragment increased; siEZH2 inhibited HepG2 cell proliferation and promote apoptosis, and led to reduced expression of Caspase-3 full-length and increased expression of active fragments. Luciferase results indicate that miR-106b does not directly target EZH2, and miR-106b promotes EZH2 expression. Conclusion Inhibition of miR-106b and EZH2 expression can effectively inhibit HepG2 cell proliferation and promote apoptosis.