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  • Original Paper
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Activation of diacylglycerol kinase α is required for VEGF-induced angiogenic signaling in vitro

Oncogene volume 23, pages 4828–4838 (2004)Cite this article

Abstract

Vascular endothelial growth factor-A (VEGF-A) promotes angiogenesis by stimulating migration, proliferation and organization of endothelium, through the activation of signaling pathways involving Src tyrosine kinase. As we had previously shown that Src-mediated activation of diacylglycerol kinase-α (Dgk-α) is required for hepatocytes growth factor-stimulated cell migration, we asked whether Dgk-α is involved in the transduction of angiogenic signaling. In PAE-KDR cells, an endothelial-derived cell line expressing VEGFR-2, VEGF-A165, stimulates the enzymatic activity of Dgk-α: activation is inhibited by R59949, an isoform-specific Dgk inhibitor, and is dependent on Src tyrosine kinase, with which Dgk-α forms a complex. Conversely in HUVEC, VEGF-A165-induced activation of Dgk is only partially sensitive to R59949, suggesting that also other isoforms may be activated, albeit still dependent on Src tyrosine kinase. Specific inhibition of Dgk-α, obtained in both cells by R59949 and in PAE-KDR by expression of Dgk-α dominant-negative mutant, impairs VEGF-A165-dependent chemotaxis, proliferation and in vitro angiogenesis. In addition, in HUVEC, specific downregulation of Dgk-α by siRNA impairs in vitro angiogenesis on matrigel, further suggesting the requirement for Dgk-α in angiogenic signaling in HUVEC. Thus, we propose that activation of Dgk-α generates a signal essential for both proliferative and migratory response to VEGF-A165, suggesting that it may constitute a novel pharmacological target for angiogenesis control.

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Acknowledgements

This study was supported by grants from University A Avogadro of Piemonte Orientale, Regione Piemonte, the Italian Ministry for University and Research (PRIN 2002-03 University research program and FIRB post-genomic program), Fondazione Cariplo (to AG), Associazione Italiana per la Ricerca sul Cancro (to AG and FB) and Istituto Superiore di Sanità (IV Programma Nazionale di Ricerca sull'AIDS-2001 to FB). GB and SC were supported by a fellowship from FIRC.

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Author notes

  1. Gianluca Baldanzi, Stefania Mitola, Federico Bussolino and Andrea Graziani: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Medical Sciences, University Amedeo Avogadro of Piemonte Orientale, v. Solaroli 17, Novara, 28100, Italy

    Gianluca Baldanzi, Santina Cutrupi, Nicoletta Filigheddu, Fabiola Sinigaglia & Andrea Graziani

  2. Institute for Cancer Research and Treatment (IRCC) and Department of Oncological Sciences, University of Torino, Italy

    Stefania Mitola & Federico Bussolino

  3. Division of Cellular Biochemistry at the Netherlands Cancer Institute, 1066 CX, Amsterdam, The Netherlands

    Wim J van Blitterswijk

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  1. Gianluca Baldanzi
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  2. Stefania Mitola
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Correspondence to Andrea Graziani.

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Baldanzi, G., Mitola, S., Cutrupi, S. et al. Activation of diacylglycerol kinase α is required for VEGF-induced angiogenic signaling in vitro. Oncogene 23, 4828–4838 (2004). https://doi.org/10.1038/sj.onc.1207633

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