Effect and mechanism of miR-155 on proliferation of laryngeal squamous cell carcinoma Hep2 cell line
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摘要: 目的 探讨miR-155对喉鳞状细胞癌Hep2细胞增殖影响及具体机制。方法 采用miR-155 NC和miR-155 inhibitor转染Hep2细胞,realtime-PCR法检测转染效果,细胞计数盒-8(CCK-8)法检测不同转染时间(12、24、36、48、60、72 h)细胞活力,流式细胞术检测细胞凋亡及细胞周期;Western blot检测Hep2细胞中细胞周期蛋白A1(cyclin A1)、cyclin D1、磷脂酰肌醇3激酶/蛋白激酶B/叉形头转录因子(Foxo3a)信号通路相关蛋白表达量。结果 miR-155 inhibitor组中miR-155表达量(0.34±0.03)明显低于miR-155 NC组(1.25±0.10),且转染24、36、48、60、72 h后,miR-155 inhibitor组Hep2细胞活力明显低于miR-155 NC组;与miR-155 NC组比较,miR-155 inhibitor组Hep2细胞凋亡率明显提高,细胞周期阻滞于G1期,cyclin A1、cyclin D1、PI3K、p-AKT表达下调,Foxo3a表达上调,差异均有统计学意义(均P<0.01)。结论 miR-155表达下调可降低Hep2细胞活力、增加细胞凋亡率,其机制可能与miR-155调控PI3K/AKT/Foxo3a信号通路有关。Abstract: Objective To explore effect and mechanism of microRNA-155 (miR-155) on the proliferation of laryngeal squamous cell carcinoma Hep2 cells.Methods Hepatocellular carcinoma Hep2 cells were transfected with miR-155 NC and miR-155 inhibitor with liposomes.The expression of miR-155 was detected with realtime-PCR.The viability of the cells with different transfection time (12,24,36,48,60,and 72 hours) was detected with Cell Counting Kit-8 (CCK-8) assay.Cell apoptosis and cell cycle were determined with flow cytometry.The expressions of cell cycle protein A1 (cyclinA1),cell cycle protein D1 (cyclinD1),phosphatidylinositol 3 kinase (PI3K)/protein kinase B (AKT),and forkhead box 3a (Foxo3a) signal pathway related proteins were detected with Western blot.Results The expression of miR-155 in the Hep2 cells transfected with miR-155 inhibitor was obviously lower than that in the cells transfected with miR-155 NC (0.34±0.03 vs.1.25±0.10) and the viability of Hep2 cells transfected with miR-155 inhibitor was lower than that of cells with miR-155 NC 24,36,48,60,and 72 hours after the transfection.Compared to those transfected with miR-155 NC,the Hep2 cells transfected with miR-155 inhibitor showed significantly increased apoptotic rate,the cell cycle arrested in G1,significantly down-regulated expressions of cyclinA1,cyclinD1,PI3K and p-AKT,and significantly up-regualated expression of Foxo3a (all P<0.01).Conclusion miR-155 inhibitor could affect the proliferation and apoptosis of Hep2 cells and the effects may relate to the regulation of PI3K/AKT/Foxo3a signal pathway.
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Key words:
- microRNA-155 /
- laryngeal squamous cell carcinoma /
- Hep2 cell /
- proliferation /
- apoptosis
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