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J. Biol. Chem., Vol. 280, Issue 23, 22454-22461, June 10, 2005

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Characterization of Heparin Affin Regulatory Peptide Signaling in Human Endothelial Cells^*

Apostolos Polykratis¶, Panagiotis Katsoris, José Courty||, and Evangelia Papadimitriou**

From the Laboratory of Molecular Pharmacology, Department of Pharmacy, and the Division of Genetics, Cell and Developmental Biology, Department of Biology, University of Patras, 26504 Patras, Greece, and the ^||Laboratoire de la Recherche sur la Croissance Cellulaire, la Reparation et la Regeneration Tissulaires, CNRS UMR 7149, Université Paris Val de Marne, Avenue du Général de Gaulle, 94010 Créteil, France

Heparin affin regulatory peptide (HARP) is an 18-kDa secreted growth factor that has a high affinity for heparin and a potent role on tumor growth and angiogenesis. We have previously reported that HARP is mitogenic for different types of endothelial cells and also affects cell migration and differentiation (12). In this study we examined the signaling pathways involved in the migration and tube formation on matrigel of human umbilical vein endothelial cells (HUVEC) induced by HARP. We report for the first time that receptor-type protein-tyrosine phosphatase / (RPTP/), which is a receptor for HARP in neuronal cell types, is also expressed in HUVEC. We also document that HARP signaling through RPTP/ leads to activation of Src kinase, focal adhesion kinase, phosphatidylinositol 3-kinase, and Erk1/2. Sodium orthovanadate, chondroitin sulfate-C, PP1, wortmannin, LY294002, and U0126 inhibit HARP-mediated signaling and HUVEC migration and tube formation. In addition, RPTP/ suppression using small interfering RNA technology interrupts intracellular signals and HUVEC migration and tube formation induced by HARP. These results establish the role of RPTP/ as a receptor of HARP in HUVEC and elucidate the HARP signaling pathway in endothelial cells.

Received for publication, December 22, 2004 , and in revised form, March 24, 2005.

^* This work was supported by grants from the Research Committee of the University of Patras (Karatheodoris) and Empeirikio Foundation, Greece. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^¶ A recipient of a fellowship from the State Scholarships Foundation.

^** To whom correspondence should be addressed. Tel.: 30-2610-969336; Fax: 30-2610-997665; E-mail: epapad{at}upatras.gr.

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