Regulation of Invasive Behavior by Vascular Endothelial Growth Factor Is HEF1-dependent

Oncogene. 2010 Aug 5;29(31):4449-59. doi: 10.1038/onc.2010.185. Epub 2010 May 24.


We previously reported a vascular endothelial growth factor (VEGF) autocrine loop in head and neck squamous cell carcinoma (HNSCC) cell lines, supporting a role for VEGF in HNSCC tumorigenesis. Using a phosphotyrosine proteomics approach, we screened the HNSCC cell line, squamous cell carcinoma-9 for effectors of VEGFR2 signaling. A cluster of proteins involved in cell migration and invasion, including the p130Cas paralog, human enhancer of filamentation 1 (HEF1/Cas-L/Nedd9) was identified. HEF1 silencing and overexpression studies revealed a role for VEGF in regulating cell migration, invasion and matrix metalloproteinase (MMP) expression in a HEF1-dependent manner. Moreover, cells plated on extracellular matrix-coated coverslips showed enhanced invadopodia formation in response to VEGF that was HEF1-dependent. Immunolocalization revealed that HEF1 colocalized to invadopodia with MT1-MMP. Analysis of HNSCC tissue microarrays for HEF1 immunoreactivity revealed a 6.5-fold increase in the odds of having a metastasis with a high HEF1 score compared with a low HEF1 score. These findings suggest that HEF1 may be prognostic for advanced stage HNSCC. They also show for the first time that HEF1 is required for VEGF-mediated HNSCC cell migration and invasion, consistent with HEF1's recent identification as a metastatic regulator. These results support a strategy targeting VEGF:VEGFR2 in HNSCC therapeutics.

Publication types

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

MeSH terms

  • Adaptor Proteins, Signal Transducing / analysis
  • Adaptor Proteins, Signal Transducing / metabolism
  • Adaptor Proteins, Signal Transducing / physiology*
  • Amino Acid Sequence
  • Carcinoma, Squamous Cell / metabolism
  • Carcinoma, Squamous Cell / pathology*
  • Case-Control Studies
  • Cell Movement / drug effects*
  • Chromatography, Liquid
  • Head and Neck Neoplasms / metabolism
  • Head and Neck Neoplasms / pathology*
  • Humans
  • Molecular Sequence Data
  • Neoplasm Invasiveness
  • Phosphoproteins / analysis
  • Phosphoproteins / metabolism
  • Phosphoproteins / physiology*
  • Proteomics
  • Pseudopodia / drug effects
  • Pseudopodia / metabolism
  • Tandem Mass Spectrometry
  • Tissue Array Analysis
  • Transfection
  • Tumor Cells, Cultured
  • Vascular Endothelial Growth Factor A / pharmacology*


  • Adaptor Proteins, Signal Transducing
  • NEDD9 protein, human
  • Phosphoproteins
  • Vascular Endothelial Growth Factor A