miR-214对人舌鳞状细胞癌增殖能力影响的体外实验研究

Effect of miR-214 on proliferation of human tongue cancer cell line in-vitro

目的 研究microRNA-214（miR-214）对舌癌AW13516细胞增殖能力的影响及其相关机制.方法 建立MIR（miR-214 mimics）组、NC（阴性转染）组及无转染对照（MOCK）组细胞.采用RT-PCR法检测各组miR-214水平;Western印迹法检测HM1、PCNA、cyclin D1的表达,采用MTT实验检测细胞增殖能力;采用软琼脂集落形成实验和平板克隆形成实验检测细胞形成克隆的能力;采用流式细胞术观察各组细胞凋亡比例变化.结果 与阴性转染组和无转染对照组相比,miR-214 mimics干预AW13516细胞后,MIR组miR-214表达水平明显升高（P＜0.05）,PIMl、PCNA、cyclin D1蛋白表达水平降低,细胞增殖能力明显降低,克隆形成能力明显减弱,凋亡比例明显升高（P＜0.05）.结论 miR-214可能通过降低PIM1水平下调PCNA、cyclin D1蛋白表达,抑制舌癌AW13516细胞的增殖能力并促进其凋亡.

Objective To investigate the effect and related mechanism ofmicroRNA-214 （miR-214） on cell proliferation of human tongue cancer cell line AW13516.Methods Cell of MIR （miR-214 mimics） group,NC （negative control mimic） group and the negative control （MOCK） group were established.The expressions of miR-214 in different groups were detected with RT-PCR,and the expressions of PIM1,PCNA,cyclin D1 with Western blot assay.MTT assay was used for study of cell proliferation.Double soft agar clone formation and plate cloning formation assay were used to measure the ability of cell cloning.The change in cell apoptosis rate was studied with flow cytometery.Results miR-214 expression of AW13516 cells transfected with miR-214 mimics was significantly up regulated at the RNA level compared with that in normal AW13516 cells （P ＜ 0.05）.Moreover,cells of MIR group showed reduced levels of PIM1,PCNA and cyclin D1 protein,remarkably lower cell proliferation,declined cloning ability and significantly elevated apoptosis rate （P＜0.05）.Conclusion miR-214 may inhibit the proliferation of tongue cancer AW13516 cells and promote their apoptosis by lowering PIM1 level and down-regulating the expressions of PCNA and cyclin D1 proteins.