Article
PDGF receptor is required for uPA-induced signalling, migration and proliferation of human vascular smooth muscle cells
Die uPA-induzierte Migration von VSMC wird durch den PDgF-Rezeptor vermittelt
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Published: | November 11, 2004 |
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In human vascular smooth muscle cell (VSMC), the Jak/Stat signaling pathway is considered as the main signaling cascade activated in response to uPA with functional consequences for cell migration and proliferation. We demonstrated recently that the Tyk2-PI3-K-RhoA-Rho kinase pathway is required for VSMC migration, whereas activation and nuclear translocation of STAT1 mediates uPA-dependent changes of VSMC proliferation. However, it remained unknown how these two cascades are linked to uPAR and which molecules serve for a downstream bifurcation of both pathways. Here we provide evidence that in VSMC platelet-derived growth factor receptor (PDGFR) serves as a transmembrane adaptor for uPAR.
We demonstrate that PDGFR tyrosine kinase activity is required for uPA to promote both, promigratory and profilerative effects. Thus, inhibition of PDGFR tyrosine kinase activity with specific inhibitor AG 1295 dose-dependently inhibited uPA-induced VSMC migration and proliferation. Further, we demonstrate that uPAR and PDGFR in VSMC are associated in a uPA-dependent manner. Thus, uPAR co-immunoprecipitated time-dependently with PDGFR from lysates of uPA stimulated VSMC but not from unstimulated cells. Similar results were obtained by co-immunoprecipitating PDGFR using anti-uPAR antibodies. PDGFR tyrosine kinase is required also for the uPA-induced STAT1 tyrosine and serine phosphorylation,as observed in Western blots. It is likely that PDGFR is capable to phosphorylate STAT1 directly, as far as in immunoprecipitation experiments we found STAT1 to be uPA-dependently associated with PDGFR in VSMC. Addition of uPA (10nM) decreased the PDGF(5ng/ml)-induced VSMC migration in Boyden chamber thus suggesting that uPAR can concure for PDGFR in order to induce uPA-dependent signaling.
Taken together, these findings suggest that PDGFR is involved in regulation of uPA-induced signaling and functional behaviour of human VSMC and can serve as a transmembrane adaptor of uPAR.