Hypoxia increases VEGF-A production by prostate cancer and bone marrow stromal cells and initiates paracrine activation of bone marrow endothelial cells

Clin Exp Metastasis. 2006;23(1):75-86. doi: 10.1007/s10585-006-9021-2. Epub 2006 Jul 7.


Hypoxia develops at sites of rapid cancer growth near sites of poorly organized vasculature. Heparin binding growth factors (HBGFs) support neoangiogenesis of tumors. We examined the effect of culturing bone-targeted, metastatic C4-2B prostate cancer cells and bone stromal derived HS27a cells under hypoxic conditions on expression of vascular endothelial growth factor (VEGF) family members. A sealed chamber infused with 1% (hypoxic) or 20% (normoxic) O(2) was used. Both cell lines produced VEGF-A in normoxia, but little or no HB-EGF, another HBGF. HS27a cells produced low levels of FGF-2 and HGF, but little or none was secreted by C4-2B cells. Levels of VEGF-A in conditioned medium (CM) from both cell lines doubled when cultured in hypoxia. Similar changes in VEGF-A mRNA levels were seen. Receptor expression was unchanged by hypoxia. Changes in VEGF-A expression during hypoxia were preceded by nuclear accumulation of hypoxia inducible factor-1alpha (HIF-1alpha). Bone marrow endothelial (BME) cells express high levels of VEGFR2/flk-1, and are targets of VEGF-A induced neovascularization. BME cells proliferated in response to treatment with HS27a CM, but not C4-2B CM. BME cells formed tube-like angiogenic structures on growth factor reduced Matrigel in response to CM from HS27a or C4-2B cells. This response was greater when CM was produced under hypoxia, and was reduced by VEGF-A or FGF-2 neutralizing antibodies. We conclude that hypoxia triggers a physiologically relevant increase in VEGF-A by prostate cancer and bone marrow stromal cells which involves a paracrine loop that recruits and activates BME to support tumor neovascularization-related processes.

Publication types

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

MeSH terms

  • Bone Marrow Cells / pathology*
  • Bone Neoplasms / secondary
  • Cell Division
  • Cell Hypoxia / physiology*
  • Cell Line, Tumor
  • DNA Primers
  • Endothelial Cells / cytology*
  • Endothelial Cells / physiology*
  • Humans
  • Male
  • Prostatic Neoplasms / genetics*
  • Prostatic Neoplasms / pathology
  • RNA, Messenger / genetics
  • RNA, Neoplasm / genetics
  • RNA, Neoplasm / isolation & purification
  • Reverse Transcriptase Polymerase Chain Reaction
  • Stromal Cells / pathology
  • Vascular Endothelial Growth Factor A / genetics*


  • DNA Primers
  • RNA, Messenger
  • RNA, Neoplasm
  • Vascular Endothelial Growth Factor A