miR-519d对绒毛膜滋养细胞增殖、侵袭、迁移能力的影响及机制

Effects and mechanism of miR-519d on proliferation,invasion and migration of human chorionic trophoblast cells

目的观察miR-519d对绒毛膜滋养细胞增殖、侵袭及迁移能力的影响,并探讨可能的作用机制。方法体外培养人绒毛膜滋养细胞HTR-8/SVneo,将HTR-8/SVneo细胞分为实验组和对照组,分别转染miR-519d mimics和NC-mimics,采用CCK-8实验检测两组细胞增殖能力,Transwell实验检测细胞侵袭、迁移能力,采用Westernblotting法检测细胞中基质金属蛋白酶2(MMP-2)蛋白。应用双荧光素酶报告基因实验验证miR-519d与MMP-2的靶向关系。结果实验组细胞增殖、侵袭、迁移能力均低于对照组,MMP-2蛋白相对表达量低于对照组(P均<0.05)。双荧光素酶报告基因证实,miR-519d能够靶向结合MMP-2的3'非编码区,MMP-2是miR-519d的靶基因。结论miR-519d高表达能够抑制绒毛膜滋养细胞增殖、侵袭、迁移能力,其机制可能与调控MMP-2表达有关。

Objective To observe the effects of miR-519d on the proliferation,invasion and migration of human cho⁃rionic trophoblast cells,and to explore the possible mechanism.Methods Human chorionic trophoblast cells HTR-8/SVneo were cultured in vitro,and were divided into the experimental group and control group,which were transfected with miR-519d mimics and NC-mimics,respectively.CCK-8 experiment was used to detect the cell proliferation ability of the two groups,Transwell experiment was used to detect the cell invasion and migration abilities,and Western blotting was used to detect the matrix metalloproteinase 2(MMP-2)protein in the cells.The targeted relationship between miR-519d and MMP-2 was verified by dual luciferase reporter gene assay.Results The cell proliferation,invasion and migration abilities in the experimental group were lower than those in the control group(all P<0.05),and the relative expression of MMP-2 protein in the experimental group was lower that in the control group(P<0.05).Dual luciferase reporter gene con⁃firmed that miR-519d could target the 3’untranslated region of MMP-2,and MMP-2 was the target gene of miR-519d.Conclusion High expression of miR-519d can inhibit the proliferation,invasion and migration of human chorionic tro⁃phoblast cells,and its mechanism may be related to the regulation of MMP-2 expression.