Regulation of invasive behavior by vascular endothelial growth factor is HEF1-dependent.
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.
Lucas
J T
JT
Department of Cell and Molecular Pharmacology and Experimental Therapeutics, Medical University of South Carolina, Charleston, SC, USA.
Salimath
B P
BP
Slomiany
M G
MG
Rosenzweig
S A
SA
D048868:Adaptor Proteins, Signal Transducing / Q000032:analysis / Q000378:metabolism / Q000502:physiology*; D000595:Amino Acid Sequence; D002294:Carcinoma, Squamous Cell / Q000378:metabolism / Q000473:pathology*; D016022:Case-Control Studies; D002465:Cell Movement / Q000187:drug effects*; D002853:Chromatography, Liquid; D006258:Head and Neck Neoplasms / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D008969:Molecular Sequence Data; D009361:Neoplasm Invasiveness; D010750:Phosphoproteins / Q000032:analysis / Q000378:metabolism / Q000502:physiology*; D040901:Proteomics; D011554:Pseudopodia / Q000187:drug effects / Q000378:metabolism; D053719:Tandem Mass Spectrometry; D046888:Tissue Array Analysis; D014162:Transfection; D014407:Tumor Cells, Cultured; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology*
D048868:Adaptor Proteins, Signal Transducing; C099942:NEDD9 protein, human; D010750:Phosphoproteins; D042461:Vascular Endothelial Growth Factor A