Targeting the cross-talk between Urokinase receptor and Formyl peptide receptor type 1 to prevent invasion and trans-endothelial migration of melanoma cells

Background Accumulating evidence demonstrates that the Urokinase Receptor (uPAR) regulates tumor cell migration through its assembly in composite regulatory units with transmembrane receptors, and $ uPAR_{88–92} $ is the minimal sequence required to induce cell motility through the Formyl Peptide Re...
Ausführliche Beschreibung

Background Accumulating evidence demonstrates that the Urokinase Receptor (uPAR) regulates tumor cell migration through its assembly in composite regulatory units with transmembrane receptors, and $ uPAR_{88–92} $ is the minimal sequence required to induce cell motility through the Formyl Peptide Receptor type 1 (FPR1). Both uPAR and FPR1 are involved in melanoma tumor progression, suggesting that they may be targeted for therapeutic purposes. In this study, the role of the uPAR-FPR1 cross-talk to sustain melanoma cell ability to invade extracellular matrix and cross endothelial barriers is investigated. Also, the possibility that inhibition of the uPAR mediated FPR1-dependent signaling may prevent matrix invasion and transendothelial migration of melanoma cells was investigated. Methods Expression levels of uPAR and FPR1 were assessed by immunocytochemistry, Western Blot and qRT-PCR. Cell migration was investigated by Boyden chamber and wound-healing assays. Migration and invasion kinetics, trans-endothelial migration and proliferation of melanoma cells were monitored in real time using the xCELLigence technology. The agonist-triggered FPR1 internalization was visualized by confocal microscope. Cell adhesion to endothelium was determined by fluorometer measurement of cell-associated fluorescence or identified on multiple z-series by laser confocal microscopy. The 3D–organotypic models were set up by seeding melanoma cells onto collagen I matrices embedded dermal fibroblasts. Data were analyzed by one-way ANOVA and post-hoc Dunnett t-test for multiple comparisons. Results We found that the co-expression of uPAR and FPR1 confers to A375 and M14 melanoma cells a clear-cut capability to move towards chemotactic gradients, to cross extracellular matrix and endothelial monolayers. FPR1 activity is required, as cell migration and invasion were abrogated by receptor desensitization. Finally, melanoma cell ability to move toward chemotactic gradients, invade matrigel or fibroblast-embedded collagen matrices and cross endothelial monolayers are prevented by anti-$ uPAR_{84–95} $ antibodies or by the RI-3 peptide which we have previously shown to inhibit the $ uPAR_{84–95} $/FPR1 interaction. Conclusions Collectively, our findings identify uPAR and FPR1 as relevant effectors of melanoma cell invasiveness and suggest that inhibitors of the $ uPAR_{84–95} $/FPR1 cross-talk may be useful for the treatment of metastatic melanoma. Ausführliche Beschreibung

Autor*in:	Ragone, Concetta [verfasserIn] Minopoli, Michele Ingangi, Vincenzo Botti, Giovanni Fratangelo, Federica Pessi, Antonello Stoppelli, Maria Patrizia Ascierto, Paolo Antonio Ciliberto, Gennaro Motti, Maria Letizia Carriero, Maria Vincenza

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2017

Schlagwörter:	Urokinase receptor Formyl peptide receptor type 1 Melanoma Peptides Trans-endothelial migration

Anmerkung:	© The Author(s). 2017

Übergeordnetes Werk:	Enthalten in: Journal of experimental & clinical cancer research - Berlin : Springer, 2008, 36(2017), 1 vom: 08. Dez.
Übergeordnetes Werk:	volume:36 ; year:2017 ; number:1 ; day:08 ; month:12

Links:	Volltext

DOI / URN:	10.1186/s13046-017-0650-x

Katalog-ID:	SPR029640954