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。
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.
作者
周芝熠
李晓娟
蒋国庆
王珏
钱源
ZHOU Zhiyi;LI Xiaojuan;JIANG Guoqing;WANG Jue;QIAN Yuan(Department of Obstetrics, First Affiliated Hospital of Kunming Medical University, Kunming 650032, China;Clinical Laboratory, First Affiliated Hospital of Kunming Medical University, Kunming 650032, China;Yunnan Provincial Key Laboratory of Laboratory Medicine, Kunming 650032, China;Yunnan Provincial Institute of Laboratory Diagnosis, Kunming 650032, China)
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2019年第4期437-442,共6页
Journal of Southern Medical University
基金
国家自然科学基金(81360103
81760273)
云南省高层次卫生技术人才培养(后备人才)(H201628)
云南省教育厅科学研究基金项目(2016ZZX090)
2017年云南省应用基础研究计划面上项目(2017FB107)~~
