Impact of IGF-I/IGF-IR Circuit on the Angiogenetic Properties of Ewing’s Sarcoma Cells

R. Strammiello; S. Benini; M. C. Manara; S. Perdichizzi; M. Serra; E. Spisni; P. Picci; K. Scotlandi

doi:10.1055/s-2004-814149

Hormone and Metabolic Research, Inhaltsverzeichnis

Horm Metab Res 2003; 35(11/12): 675-684
DOI: 10.1055/s-2004-814149

Original

Impact of IGF-I/IGF-IR Circuit on the Angiogenetic Properties of Ewing’s Sarcoma Cells

R. Strammiello ¹ , S. Benini ¹ , M. C. Manara ¹ , S. Perdichizzi ¹ , M. Serra ¹ , E. Spisni ² , P. Picci ¹ , K. Scotlandi ¹

¹Laboratorio di Ricerca Oncologica, Istituti Ortopedici Rizzoli, Bologna, Italy
²Dept. of Experimental Biology, University of Bologna, Bologna, Italy

Abstract

The insulin-like growth factor receptor I (IGF-I)-mediated circuit is a major autocrine loop for Ewing’s sarcoma (ES) cells, and plays a role the pathogenesis and malignancy of this tumor. IGF-I receptor (IGF-IR) has emerged as a good therapeutic site for ES patients. In this study, we analyzed the impact of strategies targeting the IGF-IR on the regulation of VEGFs, which are of fundamental importance in angiogenesis, and TGFβ, CTGF and Cyr61, which are factors primarily involved in skeletal growth control and angiogenesis. IGF-I increases expression of VEGF-A, TGFβ, CTGF and Cyr61 mRNA. However, only the modulation of VEGF-A expression appears to be mediated by IGF-IR. Functional assays on endothelial cells indicate a strict correlation between survival and proliferation of HUVECs and VEGF-A levels, confirming a major role for this factor in angiogenesis. Blockage of IGF-IR functions by neutralizing antibody or antisense strategies significantly reduced the expression and secretion of VEGF-A by ES cells, and supernatants of treated cells were unable to sustain the survival and proliferation of HUVECs. Analysis of the signaling mechanisms involved in constitutive or IGF-induced expression and secretion of VEGF-A indicated that PI3-K and MAPK signaling pathways are both required for VEGF expression and production in ES cells. Selective inhibitors LY294002 or PD98059 were highly effective in reducing the ability of ES cells to produce VEGF-A and stimulate survival and proliferation of HUVECs. Taken together, these findings add a new activity to the IGF-I repertoire in ES and highlight how disruption of IGF-IR functions may constitute an effective tool for the control of neovascularization in this tumor.

Key words

IGF-I - VEGF - Ewing’s sarcoma - CTGF - Cyr61 - TGFβ

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