Vascular endothelial growth factor receptor-3 and focal adhesion kinase bind and suppress apoptosis in breast cancer cells.
Focal adhesion kinase (FAK) and vascular endothelial growth factor receptor-3 (VEGFR-3) are protein tyrosine kinases that are overexpressed in human cancer and play an important role in survival signaling. In addition to its involvement with cell survival, VEGFR-3 is a primary factor in lymphatic angiogenesis. Because FAK function is regulated by its COOH terminus (FAK-CD), we used FAK-CD as a target to identify binding partners. We isolated a peptide from a phage library that bound to FAK-CD, specifically the focal adhesion targeting domain of FAK and was homologous to VEGFR-3, suggesting these two tyrosine kinases physically interact. We have also shown that VEGFR-3 is overexpressed in human breast tumors and cancer cell lines. For the first time, we have shown the physical association of FAK and VEGFR-3. The association between the NH(2) terminus of VEGFR-3, containing the peptide identified by phage display, and the COOH terminus of FAK was detected by in vitro and in vivo binding studies. We then coupled a 12-amino-acid VEGFR-3 peptide, AV3, to a TAT cellular penetration sequence and showed that AV3 and not control-scrambled peptide caused specific displacement of FAK from the focal adhesions and affected colocalization of FAK and VEGFR-3. In addition, AV3 peptide decreased proliferation and caused cell detachment and apoptosis in breast cancer cell lines but not in normal breast cells. Thus, the FAK/VEGFR-3 interaction may have a potential use to develop novel molecular therapeutics to target the signaling between FAK and VEGFR-3 in human tumors.
Garces
Christopher A
CA
Department of Surgery, Biochemistry, and Molecular Biology, University of Florida, PO Box 100286, 1600 Southwest Archer Road, Gainesville, FL 32610, USA.
Kurenova
Elena V
EV
Golubovskaya
Vita M
VM
Cance
William G
WG
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D001667:Binding, Competitive; D001943:Breast Neoplasms / Q000201:enzymology* / Q000473:pathology; D002448:Cell Adhesion / Q000502:physiology; D048708:Cell Growth Processes / Q000502:physiology; D045744:Cell Line, Tumor; D003593:Cytoplasm / Q000201:enzymology; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism*; D006801:Humans; D051379:Mice; D010446:Peptide Fragments / Q000378:metabolism; D011485:Protein Binding; D040321:Vascular Endothelial Growth Factor Receptor-3 / Q000096:biosynthesis / Q000378:metabolism*
D010446:Peptide Fragments; D040321:Vascular Endothelial Growth Factor Receptor-3; D051416:Focal Adhesion Protein-Tyrosine Kinases