Integrin β3 Mediates the Endothelial-to-Mesenchymal Transition via the Notch Pathway

Cell Physiol Biochem. 2018;49(3):985. doi: 10.1159/000493229. Epub 2018 Sep 7.

Abstract

Background/aims: Neointimal hyperplasia is responsible for stenosis, which requires corrective vascular surgery, and is also a major morphological feature of many cardiovascular diseases. This hyperplasia involves the endothelial-to-mesenchymal transition (EndMT). We investigated whether integrin β3 can modulate the EndMT, as well as its underlying mechanism.

Methods: Integrin β3 was overexpressed or knocked down in human umbilical vein endothelial cells (HUVECs). The expression of endothelial markers and mesenchymal markers was determined by real-time reverse transcription PCR (RT-PCR), immunofluorescence staining, and western blot analysis. Notch signaling pathway components were detected by real-time RT-PCR and western blot analysis. Cell mobility was evaluated by wound-healing, Transwell, and spreading assays. Fibroblast-specific protein 1 (FSP-1) promoter activity was determined by luciferase assay.

Results: Transforming growth factor (TGF)-β1 treatment or integrin β3 overexpression significantly promoted the EndMT by downregulating VE-cadherin and CD31 and upregulating smooth muscle actin α and FSP-1 in HUVECs, and by enhancing cell migration. Knockdown of integrin β3 reversed these effects. Notch signaling was activated after TGF-β1 treatment of HUVECs. Knockdown of integrin β3 suppressed TGF-β1-induced Notch activation and expression of the Notch downstream target FSP-1.

Conclusion: Integrin β3 may promote the EndMT in HUVECs through activation of the Notch signaling pathway.

Keywords: Integrin β3; Notch signaling pathway; endothelial cells; endothelial-to-mesenchymal transition; neointimal hyperplasia.

MeSH terms

  • Actins / metabolism
  • Adult
  • Aged
  • Aged, 80 and over
  • Antigens, CD / metabolism
  • Arteriovenous Fistula / metabolism
  • Arteriovenous Fistula / pathology
  • Cadherins / metabolism
  • Calcium-Binding Proteins / metabolism
  • Cell Movement / drug effects
  • Endothelial Cells / cytology
  • Endothelial Cells / drug effects
  • Endothelial Cells / metabolism
  • Epithelial-Mesenchymal Transition* / drug effects
  • Human Umbilical Vein Endothelial Cells
  • Humans
  • Integrin beta3 / chemistry
  • Integrin beta3 / genetics
  • Integrin beta3 / metabolism*
  • Middle Aged
  • RNA Interference
  • RNA, Small Interfering / metabolism
  • Receptors, Notch / metabolism*
  • S100 Calcium-Binding Protein A4
  • Signal Transduction* / drug effects
  • Transforming Growth Factor beta1 / pharmacology
  • Up-Regulation / drug effects

Substances

  • ACTA2 protein, human
  • Actins
  • Antigens, CD
  • Cadherins
  • Calcium-Binding Proteins
  • Integrin beta3
  • RNA, Small Interfering
  • Receptors, Notch
  • S100 Calcium-Binding Protein A4
  • Transforming Growth Factor beta1
  • cadherin 5
  • S100A4 protein, human