miR-195靶向AEG-1基因对子宫内膜癌细胞增殖、凋亡、迁移、侵袭的影响及机制研究

Effects of miR-195 targeting AEG-1 gene on proliferation,apoptosis,migration and invasion of endometrial cancer cells and its action mechanism

目的探讨miR-195对子宫内膜癌HEC-1A细胞增殖、凋亡、迁移和侵袭的影响,并初步探讨其作用机制。方法采用脂质体介导将miR-195模拟物(mimics)转染至HEC-1A细胞为miR-195组,转染阴性对照和未转染的HEC-1A细胞分别为NC组和Blank组。qPCR检测各组HEC-1A细胞中miR-195的表达水平,MTT和流式细胞术分别检测细胞增殖和凋亡情况,Transwell实验检测细胞迁移和侵袭能力,qPCR和Westernblot检测AEG-1mRNA和蛋白表达水平,双荧光素酶报告基因实验验证miR-195和HEC-1A的靶向关系。结果与Blank组和NC组比,miR-195组HEC-1A细胞中miR-195的表达水平明显上调(P<0.05),OD值明显降低(P<0.05),凋亡率明显升高(P<0.05),迁移和侵袭细胞数显著减少(P<0.05),AEG-1mRNA和蛋白表达水平明显下调(P<0.05),双荧光素酶报告基因实验结果显示,AEG-1-Wt组中miR-195组HEC-1A细胞的相对荧光活性显著低于NC组(P<0.05),AEG-1-Mut组中miR-195组HEC-1A细胞的相对荧光活性与NC组比较差异无统计学意义(P>0.05)。结论 miR-195能够抑制子宫内膜癌HEC-1A细胞增殖、迁移和侵袭,并促进HEC-1A细胞凋亡,该过程可能与靶向调控AEG-1的表达有关。

Objective To investigate the effects of miR-195 on proliferation,apoptosis,migration and invasion of endometrial carcinoma HEC-1 A cells,and to explore its action mechanism.Methods The miR-195 mimics were transfected into HEC-1 A cells by liposome(miR-195 group).The transfected negative control and untransfected HEC-1 A cells were used as NC group and Blank group,respectively.The qPCR was used to detect the expression levels of miR-195 in HEC-1 A cells.MTT and flow cytometry were used to detect cell proliferation and apoptosis,and Transwell assay was used to detect cell migration and invasion,moreover,the expression levels AEG-1 mRNA and protein were detected by qPCR and Western blot.In addition the targeting correlation between between miR-195 and HEC-1 A was identified by dual luciferase reporter assays.Results As compared with those in blank group and NC group,the expression levels of miR-195 in the HER-1 A cells were significantly up-regulated(P<0.05),the OD value was significantly decreased(P<0.05),and the apoptosis rate was significantly increased(P<0.05),the number of migration and invasion cells was significantly decreased(P<0.05),and the expression levels of AEG-1 mRNA and protein were significantly down-regulated(P<0.05). The results of dual luciferase reporter gene assay showed that in AEG-1-Wt group the relative fluorescence activity of HEC-1 A cells in miR-195 group was significantly lower than that in NC group(P<0.05).The relative fluorescence activity of HEC-1 A cells in miR-195 group in AEG-1-Mut group was not significantly different from that in NC group(P>0.05).Conclusion The miR-195 can inhibit the proliferation,migration and invasion of endometrial cancer HEC-1 A cells and promote the apoptosis of HEC-1 A cells,which may be related to the targeted regulation of AEG-1 expression.