Bone morphogenetic protein 4 regulates microRNAs miR-494 and miR-126-5p in control of endothelial cell function in angiogenesis

Thromb Haemost. 2017 Apr 3;117(4):734-749. doi: 10.1160/TH16-08-0643. Epub 2017 Jan 26.


MicroRNAs are small non-coding RNAs that negatively regulate posttranscriptional gene expression. Several microRNAs have been described to regulate the process of angiogenesis. Previously, we have shown that bone morphogenetic protein 4 (BMP4) increased the pro-angiogenic activity of endothelial cells. In this project, we now investigated how the pro-angiogenic BMP4 effect is mediated by microRNAs. First, we performed a microRNA array with BMP4-stimulated human umbilical vein endothelial cells (HUVECs). Among the top-regulated microRNAs, we detected a decreased expression of miR-494 and increased expression of miR-126-5p. Next, we analysed the canonical Smad and alternative signalling pathways, through which BMP4 would regulate miR-126-5p and miR-494 expression. Furthermore, the functional effect of miR-494 and miR-126-5p on endothelial cells was investigated. MicroRNA-494 overexpression decreased endothelial cell proliferation, migration and sprout formation. Consistently, miR-494 inhibition increased endothelial cell function. As potential miR-494 targets, bFGF and BMP endothelial cell precursor-derived regulator (BMPER) were identified and confirmed by western blot. Luciferase assays showed direct miR-494 binding in BMPER 3'UTR. In contrast, miR-126-5p overexpression increased pro-angiogenic endothelial cell behaviour and, accordingly, miR-126-5p inhibition decreased endothelial cell function. As a direct miR-126-5p target we identified the anti-angiogenic thrombospondin-1 which was confirmed by western blot analysis and luciferase assays. In the Matrigel plug assay application of antagomiR-494 increased endothelial cell ingrowth, whereas antagomiR-126-5p treatment decreased cell ingrowth in vivo. Taken together, through differential regulation of the anti-angiomiR-494 and the angiomiR-126-5p by BMP4 both microRNAs contribute to the pro-angiogenic BMP4 effect on endothelial cells.

Keywords: BMP4; angiogenesis; endothelial cells; microRNAs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3' Untranslated Regions
  • Angiogenesis Inducing Agents / pharmacology*
  • Animals
  • Binding Sites
  • Bone Morphogenetic Protein 4 / pharmacology*
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism
  • Cell Movement / drug effects
  • Cell Proliferation / drug effects
  • Fibroblast Growth Factor 2 / genetics
  • Fibroblast Growth Factor 2 / metabolism
  • Gene Expression Regulation
  • Human Umbilical Vein Endothelial Cells / drug effects*
  • Human Umbilical Vein Endothelial Cells / metabolism
  • Humans
  • Mice
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • NIH 3T3 Cells
  • Neovascularization, Physiologic / drug effects*
  • Signal Transduction / drug effects
  • Thrombospondin 1 / genetics
  • Thrombospondin 1 / metabolism
  • Transfection


  • 3' Untranslated Regions
  • Angiogenesis Inducing Agents
  • BMP4 protein, human
  • BMPER protein, human
  • Bone Morphogenetic Protein 4
  • Carrier Proteins
  • MIRN126 microRNA, human
  • MIRN494 microRNA, human
  • MicroRNAs
  • Thrombospondin 1
  • thrombospondin-1, human
  • Fibroblast Growth Factor 2