Inhibition of endothelial cell chemotaxis toward FGF-2 by gefitinib associates with downregulation of Fes activity.
Gefitinib inhibits epidermal growth factor-independent angiogenesis, but the molecular mechanism underlying this inhibition has yet to be defined. Here we show that gefitinib dose-dependently inhibited chemotaxis of endothelial cells toward fibroblast growth factor-2 (FGF-2), but not toward vascular endothelial growth factor-A (VEGF-A). Gefitinib inhibited lamellipodium formation by endothelial cells induced by FGF-2, but not by VEGF-A. Gefitinib at 10 microM did not inhibit autophosphorylation of FGF receptor 1 or VEGF receptor 2. A non-receptor protein tyrosine kinase, Fes, has two coiled-coil domains (CCDs) in its N-terminal region. Fes is activated by trans-autophosphorylation through CCD functions. An inactivating mutation in the second CCD abolished FGF-2 activation of Fes, indicating involvement of this CCD in FGF-2-induced Fes activation. Gefitinib-treatment decreased both CCD-independent and FGF-2- or VEGF-A-promoted Fes activity with a maximal decrease at 1 microM. The same results were observed in cells stably expressing kinase-inactive Fes; a dominant negative effect was observed in cells treated with FGF-2, but not with VEGF-A. Taken together, these results indicate that FGF-2 activates Fes via the second CCD, leading to lamellipodium formation and chemotaxis by endothelial cells, and gefitinib may act through Fes as an inhibitor of FGF-2-driven angiogenesis.
Kanda
Shigeru
S
Department of Experimental and Clinical Laboratory Medicine, National Hospital Organization, Nagasaki Hospital, Nagasaki 850-8523, Japan. skanda-jua@umin.net
Naba
Alexandra
A
Miyata
Yasuyoshi
Y
D000970:Antineoplastic Agents / Q000494:pharmacology*; D015153:Blotting, Western; D002633:Chemotaxis / Q000187:drug effects*; D015536:Down-Regulation; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism; D000077156:Gefitinib; D006801:Humans; D009389:Neovascularization, Pathologic / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D051578:Proto-Oncogene Proteins c-fes / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D011799:Quinazolines / Q000494:pharmacology*; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
D000970:Antineoplastic Agents; D011799:Quinazolines; D042461:Vascular Endothelial Growth Factor A; D016222:Fibroblast Growth Factor 2; D051578:Proto-Oncogene Proteins c-fes; D000077156:Gefitinib