Platelet-derived growth factor-C promotes human melanoma aggressiveness through activation of neuropilin-1.

Federica Ruffini,Lauretta Levati,Pedro M Lacal,Simona Caporali,Grazia Graziani,Stefania D’Atri,Maria Grazia Atzori

doi:10.18632/oncotarget.18706

Abstract

Despite recent progress in advanced melanoma therapy, identification of signalling pathways involved in melanoma switch from proliferative to invasive states is still crucial to uncover new therapeutic targets for improving the outcome of metastatic disease. Neuropilin-1 (NRP-1), a co-receptor for vascular endothelial growth factor-A (VEGF-A) tyrosine kinase receptors (VEGFRs), has been suggested to play a relevant role in melanoma progression. NRP-1 can be activated by VEGF-A also in the absence of VEGFRs, triggering specific signal transduction pathways (e.g. p130Cas phosphorylation). Since melanoma cells co-expressing high levels of NRP-1 and platelet derived growth factor-C (PDGF-C) show a highly invasive behaviour and PDGF-C shares homology with VEGF-A, in this study we have investigated whether PDGF-C directly interacts with NRP-1 and promotes melanoma aggressiveness. Results demonstrate that PDGF-C specifically binds in vitro to NRP-1. In melanoma cells expressing NRP-1 but lacking PDGFRα, PDGF-C stimulates extra-cellular matrix (ECM) invasion and induces p130Cas phosphorylation. Blockade of PDGF-C function by neutralizing antibodies or reduction of its secretion by specific siRNA inhibit ECM invasion and vasculogenic mimicry. Moreover, PDGF-C silencing significantly down-modulates the expression of Snail, a transcription factor involved in tumour invasiveness that is highly expressed in NRP-1 positive melanoma cells. In conclusion, our results demonstrate for the first time a direct activation of NRP-1 by PDGF-C and strongly suggest that autocrine and/or paracrine stimulation of NRP-1 by PDGF-C might contribute to the acquisition of a metastatic phenotype by melanoma cells.

Highlights

Cutaneous melanoma is an extremely aggressive skin cancer with a highly metastatic potential
vascular endothelial growth factor (VEGF)-A interacts with two high-affinity transmembrane tyrosine www.impactjournals.com/oncotarget kinase receptors (VEGFR-1 and VEGF receptors (VEGFRs)-2) [3], but it binds to neuropilin-1 (NRP-1), a transmembrane polypeptide that, acting as co-receptor, amplifies the signal transmitted by vascular endothelial growth factor-A (VEGF-A) through VEGFR-2 [4]
Conditioned medium collected from M14-N cells promoted their invasion through extra-cellular matrix (ECM) and this effect was strongly reduced by an anti-platelet-derived growth factor (PDGF)-C antibody, to a higher extent as compared with a VEGF-A neutralizing antibody (Figure 1B and 1C), which is released at high levels by M14-N cells [5]

Summary

Introduction

Cutaneous melanoma is an extremely aggressive skin cancer with a highly metastatic potential. The molecular mechanisms associated with the acquisition of a metastatic phenotype by melanoma cells are not very well defined and, the identification of signalling pathways involved in the metastatic switch is of primary importance. Melanoma progression is greatly favoured by angiogenesis and appears to be associated with an increase of vascular endothelial growth factor-A (VEGF-A) expression by tumour cells [2]. Expression of NRP-1 in melanoma cells correlates with tumour cell invasive behaviour and formation of tube-like structures resembling vascular networks (vasculogenic mimicry). These effects derive from up-regulation of VEGF-A, matrix metalloprotease (MMP-2, MMP-9) secretion [5] and activation of specific αv integrins [6]. High NRP-1 expression is involved in tumour resistance to VEGF-A blocking therapies [7], suggesting that inhibition of NRP-1 may be required to prevent compensatory escape mechanisms by tumour cells

Methods

Results

Conclusion