p53-responsive miR-194 inhibits thrombospondin-1 and promotes angiogenesis in colon cancers

Cancer Res. 2011 Dec 15;71(24):7490-501. doi: 10.1158/0008-5472.CAN-11-1124. Epub 2011 Oct 25.

Abstract

Thrombospondin-1 (TSP-1) is an endogenous inhibitor of angiogenesis encoded by the THBS1 gene, whose promoter is activated by p53. In advanced colorectal cancers (CRC), its expression is sustained or even slightly increased despite frequent loss of p53. Here, we determined that in HCT116 CRC cells, p53 activates the THBS1 primary transcript, but fails to boost THBS1 mRNA or protein levels, implying posttranscriptional regulation by microRNAs (miRNA). In a global miRNA gain-of-function screen done in the Dicer-deficient HCT116 variant, several miRNAs negatively regulated THBS1 mRNA and protein levels, one of them being miR-194. Notably, in agreement with published data, p53 upregulated miR-194 expression in THBS1 retrovirus-transduced HCT116 cells, leading to decreased TSP-1 levels. This negative effect was mediated by a single miR-194 complementary site in the THBS1 3'-untranslated region, and its elimination resulted in TSP-1 reactivation, impaired angiogenesis in Matrigel plugs, and reduced growth of HCT116 xenografts. Conversely, transient overexpression of miR-194 in HCT116/THBS1 cells boosted Matrigel angiogenesis, and its stable overexpression in Ras-induced murine colon carcinomas increased microvascular densities and vessel sizes. Although the overall contribution of miR-194 to neoplastic growth is context dependent, p53-induced activation of this GI tract-specific miRNA during ischemia could promote angiogenesis and facilitate tissue repair.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • 3' Untranslated Regions / genetics
  • Animals
  • Blotting, Western
  • Cell Line, Tumor
  • Colonic Neoplasms / blood supply
  • Colonic Neoplasms / genetics*
  • Colonic Neoplasms / pathology
  • Gene Expression Regulation, Neoplastic
  • HCT116 Cells
  • Humans
  • Mice
  • MicroRNAs / genetics*
  • MicroRNAs / metabolism
  • Mutation
  • Neovascularization, Pathologic / genetics*
  • Retroviridae / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Thrombospondin 1 / genetics*
  • Thrombospondin 1 / metabolism
  • Transcription, Genetic
  • Transduction, Genetic
  • Transplantation, Heterologous
  • Tumor Suppressor Protein p53 / genetics*
  • Tumor Suppressor Protein p53 / metabolism

Substances

  • 3' Untranslated Regions
  • MIRN194 microRNA, human
  • MicroRNAs
  • Thrombospondin 1
  • Tumor Suppressor Protein p53