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Glioblastoma-secreted factors induce IGFBP7 and angiogenesis by modulating Smad-2-dependent TGF-β signaling

Oncogene volume 27, pages 6834–6844 (2008)Cite this article

Abstract

Insulin-like growth factor-binding protein 7 (IGFBP7) is a selective biomarker of glioblastoma (GBM) vessels, strongly expressed in tumor endothelial cells and vascular basement membrane. IGFBP7 gene regulation and its potential role in tumor angiogenesis remain unclear. Mechanisms of IGFBP7 induction and its angiogenic capacity were examined in human brain endothelial cells (HBECs) exposed to tumor-like conditions. HBEC treated with GBM cell (U87MG)-conditioned media (-CM) exhibited fourfold upregulation of IGFBP7 mRNA and protein compared to control cells. IGFBP7 gene regulation in HBEC was methylation independent. U87MG-CM analysed by enzyme-linked immunosorbent assay contained ∼5 pM transforming growth factor (TGF)-β1, a concentration sufficient to stimulate IGFBP7 in HBEC to similar levels as U87MG-CM. Both pan-TGF-β-neutralizing antibody (1D11) and the TGF-β1 receptor (activin receptor-like kinase 5, ALK5) antagonist, SB431542, blocked U87MG-CM-induced IGFBP7 expression in HBEC, indicating that TGF-β1 is an important tumor-secreted effector capable of IGFBP7 induction in endothelial cells. HBEC exposed to either U87MG-CM or IGFBP7 protein exhibited increased capillary-like tube (CLT) formation in Matrigel. Both TGF-β1- and U87MG-CM-induced Smad-2 phosphorylation and U87MG-CM-induced CLT formation in HBEC were inhibited by the ALK5 antagonist, SB431542. These data suggest that proangiogenic IGFBP7 may be induced in brain endothelial cells by TGF-βs secreted by GBM, most likely through TGF-β1/ALK5/Smad-2 pathway.

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Acknowledgements

We thank Gilles St-Laurent for technical assistance for the production of IGFBP7, Mrs Debbie Callaghan for technical assistance in nuclear protein extraction, Dr Anne Lenferink for valuable discussions and Mr Tom Devecseri for image processing.

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Authors and Affiliations

  1. Cerebrovascular Research Group, Neurobiology Program, Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario, Canada

    A Pen, M J Moreno & D B Stanimirovic

  2. Department of Cellular and Molecular Medicine, Faculty of Medicine, University of Ottawa, Ottawa, Ontario, Canada

    A Pen & D B Stanimirovic

  3. Animal Cell Technology Group, Biotechnology Research Institute, National Research Council of Canada, Montreal, Quebec, Canada

    Y Durocher

  4. Neurogenesis and Brain Repair Group, Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario, Canada

    P Deb-Rinker

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Pen, A., Moreno, M., Durocher, Y. et al. Glioblastoma-secreted factors induce IGFBP7 and angiogenesis by modulating Smad-2-dependent TGF-β signaling. Oncogene 27, 6834–6844 (2008). https://doi.org/10.1038/onc.2008.287

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