ELK3 suppresses angiogenesis by inhibiting the transcriptional activity of ETS-1 on MT1-MMP

Int J Biol Sci. 2014 Mar 27;10(4):438-47. doi: 10.7150/ijbs.8095. eCollection 2014.

Abstract

Ets transcription factors play important roles in vasculogenesis and angiogenesis. Knockout of the Ets gene family members in mice resulted in disrupted angiogenesis and malformed vascular systems. In this study, the role and mechanism of ELK3, an Ets factor, in angiogenesis was investigated using ELK3-specific siRNA in human vascular endothelial cells (HUVECs) and in vivo implantation assay. The suppression of ELK3 expression resulted in the reinforcement of VEGF-induced tube formation in HUVECs. The in vivo Matrigel plug assay also showed that ELK3 knockdown resulted in increased angiogenesis. Luciferase activity of the MT1-MMP promoter induced by ETS-1 factor was attenuated ELK3 co-transfection. CHIP assay showed the binding of ELK3 on the MT1-MMP promoter. MT1-MMP knockdown in the ELK3 knockdowned cells resulted in the decrease of tube formation suggesting that MT1-MMP transcriptional repression is required for ELK3-mediated anti-angiogenesis effect. Our data also showed that the suppressive effect of ELK3 on the angiogenesis was partly due to the inhibitory effect of ELK3 to the ETS-1 transcriptional activity on the MT1-MMP promoter rather than direct suppression of ELK3 on the target gene, since the expression level of co-repressor Sin3A is low in endothelial cells. Our results suggest that ELK3 plays a negative role of VEGF-induced angiogenesis through indirectly inhibiting ETS-1 function.

Keywords: ELK3; MT1-MMP; Sin3A.; VEGF.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Endothelial Cells / metabolism
  • Gene Expression Regulation
  • Gene Knockdown Techniques
  • Human Umbilical Vein Endothelial Cells
  • Male
  • Matrix Metalloproteinase 14 / genetics*
  • Mice
  • Mice, Inbred BALB C
  • Neovascularization, Physiologic / genetics*
  • Proto-Oncogene Protein c-ets-1 / genetics*
  • Proto-Oncogene Proteins / genetics
  • Proto-Oncogene Proteins / metabolism
  • Proto-Oncogene Proteins / physiology*
  • Proto-Oncogene Proteins c-ets
  • RNA, Messenger / metabolism
  • Repressor Proteins / metabolism
  • Repressor Proteins / physiology
  • Sin3 Histone Deacetylase and Corepressor Complex
  • Transcription Factors / genetics
  • Transcription Factors / metabolism
  • Transcription Factors / physiology*
  • Vascular Endothelial Growth Factor A / pharmacology

Substances

  • ETS1 protein, human
  • Elk3 protein, human
  • Proto-Oncogene Protein c-ets-1
  • Proto-Oncogene Proteins
  • Proto-Oncogene Proteins c-ets
  • RNA, Messenger
  • Repressor Proteins
  • SIN3A transcription factor
  • Transcription Factors
  • Vascular Endothelial Growth Factor A
  • MMP14 protein, human
  • Matrix Metalloproteinase 14
  • Sin3 Histone Deacetylase and Corepressor Complex