维生素D通过下调miR-21的表达促进人胎盘滋养细胞的迁移和侵袭-《南方医科大学学报》

文章信息/Info

Title:: Vitamin D down-regulates microRNA-21 expression to promote human placental trophoblast cell migration and invasion in vitro

Keywords:: vitaminD; microRNA-21; trophoblasts; HTR-8/SVneo cells; migration; human placental trophoblast cells

摘要:: 目的探讨维生素D对microRNA-21表达的调控作用以及对人胎盘滋养细胞迁移、侵袭能力的影响。方法不同剂量维生素D刺激HTR-8/SVneo细胞24、48、72 h后，RT-qPCR 检测microRNA-21表达情况；将miR-21 mimic、miR-21 mimic-nc、miR- 21 inhibitor和miR-21 inhibitor-nc分别转染至HTR-8/SVneo细胞，或将miR-21 mimic、miR-21 inhibitor联合维生素D共同刺激细胞，Transwell实验检测细胞的迁移和侵袭能力，western blot 检测E-cadherin、Fibronectin、MMP9的蛋白表达水平。结果维生素D刺激HTR-8/SVneo细胞后抑制了micoRNA-21的表达，且抑制作用呈浓度-时间依赖性增强。单独转染miR-21 mimic抑制HTR-8/SVneo细胞的迁移、侵袭能力，E-cadherin表达水平增加，Fibronectin、MMP9表达水平降低；联合维生素D共同刺激细胞则可以减弱该作用。单独转染miR-21 inhibitor 促进了HTR-8/SVneo 细胞的迁移、侵袭能力，E-cadherin 表达水平降低， Fibronectin、MMP9表达水平增加；联合维生素D共同作用与单细胞独转染miR-21 inhibitor相比，细胞迁移、侵袭能力以及蛋白水平无明显改变。结论维生素D可能通过下调microRNA-21的表达来影响子痫前期发生发展。

Abstract:: Objective To investigate the effect of vitamin D on microRNA-21 (miR-21) expression and migration and invasion of human placental trophoblast cells. Methods The changes in the expression of miR-21 were detected using RT-qPCR in HTR-8/ SVneo cells following stimulation by vitamin D at different doses for 24, 48 and 72 h. HTR-8/SVneo cells transfected with miR-21 mimic or inhibitor with or without vitamin D treatment were examined for changes in cell migration and invasion abilities using Transwell assay, and Western blotting was used to detect protein expressions of E-cadherin, fibronectin, and MMP9. Results Vitamin D obviously inhibited the expression of micoRNA-21 in HTR-8/SVneo cells in a concentration-and time- dependent manner. Transfection with the miR-21 mimic significantly inhibited the migration and invasion of HTR-8/ SVneo cells, and this inhibitory effect was abolished by treatment with vitamin D; transfection with miR-21 inhibitor obviously promoted the migration and invasion of HTR-8/SVneo cells, and these effects were not significantly affected by vitamin D treatment. Conclusion Vitamin D may promote trophoblast cell migration and invasion to accelerate the development of preeclampsia by down-regulating the expression of miR-21.