Canonical hedgehog signaling augments tumor angiogenesis by induction of VEGF-A in stromal perivascular cells

Proc Natl Acad Sci U S A. 2011 Jun 7;108(23):9589-94. doi: 10.1073/pnas.1017945108. Epub 2011 May 19.

Abstract

Hedgehog (Hh) signaling is critical to the patterning and development of a variety of organ systems, and both ligand-dependent and ligand-independent Hh pathway activation are known to promote tumorigenesis. Recent studies have shown that in tumors promoted by Hh ligands, activation occurs within the stromal microenvironment. Testing whether ligand-driven Hh signaling promotes tumor angiogenesis, we found that Hh antagonism reduced the vascular density of Hh-producing LS180 and SW480 xenografts. In addition, ectopic expression of sonic hedgehog in low-Hh-expressing DLD-1 xenografts increased tumor vascular density, augmented angiogenesis, and was associated with canonical Hh signaling within perivascular tumor stromal cells. To better understand the molecular mechanisms underlying Hh-mediated tumor angiogenesis, we established an Hh-sensitive angiogenesis coculture assay and found that fibroblast cell lines derived from a variety of human tissues were Hh responsive and promoted angiogenesis in vitro through a secreted paracrine signal(s). Affymetrix array analyses of cultured fibroblasts identified VEGF-A, hepatocyte growth factor, and PDGF-C as candidate secreted proangiogenic factors induced by Hh stimulation. Expression studies of xenografts and angiogenesis assays using combinations of Hh and VEGF-A inhibitors showed that it is primarily Hh-induced VEGF-A that promotes angiogenesis in vitro and augments tumor-derived VEGF to promote angiogenesis in vivo.

MeSH terms

  • Animals
  • Cell Line
  • Cell Line, Tumor
  • Culture
  • Endothelial Cells / cytology
  • Endothelial Cells / metabolism
  • Female
  • Gene Expression Profiling
  • Gene Expression Regulation, Neoplastic
  • Hedgehog Proteins / genetics*
  • Mice
  • Mice, Nude
  • Myofibroblasts / cytology
  • Myofibroblasts / metabolism
  • Neoplasms / blood supply
  • Neoplasms / genetics*
  • Neoplasms / pathology
  • Neoplasms, Experimental / blood supply
  • Neoplasms, Experimental / genetics
  • Neoplasms, Experimental / pathology
  • Neovascularization, Pathologic / genetics*
  • Neovascularization, Physiologic / genetics
  • Oligonucleotide Array Sequence Analysis
  • Patched Receptors
  • Receptors, Cell Surface / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction / genetics*
  • Stromal Cells / metabolism
  • Stromal Cells / pathology
  • Transplantation, Heterologous
  • Vascular Endothelial Growth Factor A / genetics*

Substances

  • Hedgehog Proteins
  • Patched Receptors
  • Receptors, Cell Surface
  • Vascular Endothelial Growth Factor A

Associated data

  • GEO/GSE29316