miR-150在复发性流产患者绒毛组织中表达及在血管生成中的作用.

Expression of miR-150in Villus of Patients with Recurrent Miscarriage and Its Effect on Angiopoiesis

目的探讨miR-150在复发性流产(recurrent miscarriage,RM)患者绒毛组织中表达以及在血管生成中的作用。方法分别用RT-qPCR法及westem印迹检测RM及人工流产患者的绒毛组织中miR-150及血管内皮生长因子(vascular endothelial growth factor,VEGF)的表达情况;人脐静脉内皮细胞(human umbilical vein endothhelial cells,HUVECs)予以转染miR-150模拟物和miR-150抑制物,观察成管情况。结果RM患者绒毛组织中miR-150的表达显著高于对照组,而VEGF蛋白水平显著低于对照组。体外予以miR-150模拟物转染HuVEcs后,HuVEcs的miRNA-150水平显著升高,而VEGF蛋白水平显著降低;予以miRNA。150抑制物转染后,miR-150水平显著降低,而vEGF蛋白水平显著升高。成管实验表明,予以miR-150模拟物处理后,HUVECs的成管长度显著降低,而miR-150抑制物转染后HuVEcs的成管长度显著增加。结论miR-150在RM患者绒毛组织中表达上调,其通过靶向调控vEGF的表达,导致绒毛血管发育不良.从而参与RM发病过程。

Objective To investigate the expression level of miR-150in the villus of patients with recurrent miscarriage and its effect on angiopoiesis.Methods Expression levels of miR-150 and VEGF protein in the villus of patients with recurrent miscarriage and controls were detected by RT-qPCR and Western blotting,respectively.HUVECs cells were transfected by miR-150mimic and inhibitor,and Western blotting was used to detect the effect of miR-150on VEGF protein expression level,and the effect of miR-150on tube formation was detected.Finally,a dual luciferase reporter gene was used to verify the targeted regulation of miR-150on VEGF.Results The expression of miR-150was significantly higher in the villus of patients with recurrent miscarriage than in control group (P <0.05).miR-150upregulation inhibited HUVECs tube formation.Conversely, the downregulation of miR-150reversed these effects.The expression of VEGF protein was significantly lower in the villus of patients recurrent miscarriage than in the control group (P <0.05).After transfection of miR-150mimic,the expression of VEGF protein was decreased.Dual luciferase reporter assays confirmed that miR-150could target VEGF expression through direct binding to the 3'UTR of VEGF.Conclusion miR-150could regulate angiogenesis by targeting VEGF expression and might be involved in the pathogenesis of RM.