Hypoxia-inducible factor 1 upregulation of both VEGF and ANGPTL4 is required to promote the angiogenic phenotype in uveal melanoma

Oncotarget. 2016 Feb 16;7(7):7816-28. doi: 10.18632/oncotarget.6868.

Abstract

Purpose: Expression of the hypoxia-inducible factor (HIF)-1-regulated gene product, vascular endothelial growth factor (VEGF), correlates with tumor vascularity in patients with uveal melanoma (UM). While the relationship between HIF-1 and VEGF in cancer is well-studied, their relative contribution to the angiogenic phenotype in UM has not previously been interrogated. Here we evaluate the contribution of HIF-1, VEGF, and a second HIF-1-regulated gene product, angiopoietin-like 4 (ANGPTL4), to angiogenesis in UM.

Experimental design: UM cells were examined for expression of HIF-1α, VEGF, and ANGPTL4. Their contribution to the angiogenic potential of UM cells was assessed using the endothelial cell tubule formation and directed in vivo angiogenesis assays. These results were corroborated in tissue from UM animal models and in tissue from patients with UM.

Results: Inhibition of VEGF partially reduced tubule formation promoted by conditioned medium from UM cells. Inhibition of ANGPTL4, which was highly expressed in hypoxic UM cells, a UM orthotopic transplant model, a UM tumor array, and vitreous samples from UM patients, inhibited the angiogenic potential of UM cells in vitro and in vivo; this effect was additive to VEGF inhibition.

Conclusions: Targeting both ANGPTL4 and VEGF may be required for the effective inhibition of angiogenesis in UM.

Keywords: angiogenesis; angiopoietin-like 4 (ANGPTL4); choroidal melanoma; hypoxia inducible factor-1α; vascular endothelial growth factor (VEGF).

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Angiopoietin-Like Protein 4
  • Angiopoietins / genetics
  • Angiopoietins / metabolism*
  • Animals
  • Apoptosis
  • Blotting, Western
  • Case-Control Studies
  • Cell Hypoxia
  • Cell Movement*
  • Cell Proliferation
  • Cells, Cultured
  • Endothelium, Vascular / metabolism
  • Endothelium, Vascular / pathology
  • Female
  • Fluorescent Antibody Technique
  • Follow-Up Studies
  • Gene Expression Regulation, Neoplastic*
  • Humans
  • Hypoxia-Inducible Factor 1, alpha Subunit / genetics
  • Hypoxia-Inducible Factor 1, alpha Subunit / metabolism*
  • Immunoenzyme Techniques
  • Melanoma / blood supply*
  • Melanoma / metabolism
  • Melanoma / pathology
  • Mice
  • Mice, Nude
  • Neoplasm Staging
  • Neovascularization, Pathologic / metabolism
  • Neovascularization, Pathologic / pathology*
  • Phenotype
  • Prognosis
  • RNA, Messenger / genetics
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Uveal Neoplasms / blood supply*
  • Uveal Neoplasms / metabolism
  • Uveal Neoplasms / pathology
  • Vascular Endothelial Growth Factor A / genetics
  • Vascular Endothelial Growth Factor A / metabolism*
  • Xenograft Model Antitumor Assays

Substances

  • ANGPTL4 protein, human
  • Angiopoietin-Like Protein 4
  • Angiopoietins
  • HIF1A protein, human
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • RNA, Messenger
  • VEGFA protein, human
  • Vascular Endothelial Growth Factor A

Supplementary concepts

  • Uveal melanoma