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Tumoral and macrophage uPAR and MMP-9 contribute to the invasiveness of B16 murine melanoma cells

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Abstract

The aim of this study was to investigate whether tumor cells as well as tumor-associated macrophages (TAMs) contribute to the generation of protease activities essential to tumor cell invasiveness, such as matrix metalloproteinase 2 and 9 (MMP-2 and MMP-9), and the urokinase-type plasminogen activator (uPA) and uPA receptor (uPAR). We found that the enhanced invasiveness through Matrigel-coated filters of B16 murine melanoma cells stimulated with IFNγ was associated with an higher expression of uPAR and MMP-9 in these cells. Moreover, treatment with anti-MMP-9 or anti-uPAR monoclonal antibodies abrogated the increase of invasiveness in IFNγ-stimulated melanoma cells, suggesting a cooperation of uPA system and MMP-9 in cytokine-stimulated invasiveness. Invasiveness through Matrigel was also enhanced in B16 melanoma cells exposed to a medium conditioned by TAMs, represented in our experimental model by thioglycollate-elicited macrophages co-cultivated with melanoma cells. Macrophages isolated from these co-cultures were found to express higher levels of uPAR and MMP-9 compared to macrophage cultures alone, and the pro-invasive activity of the co-culture-conditioned medium was abrogated by anti-MMP-9 monoclonal antibodies, but not anti-uPAR monoclonal antibodies. Furthermore, the enhanced uPAR and MMP-9 expression in macrophages co-cultivated with tumor cells seems a rather specific phenomenon, generated through a cell-to-cell contact mechanism. On the whole, our data point to a cooperation between tumor cells and macrophages elicited by tumor cells themselves in generating key enzymes essential in the promotion of tumor invasiveness, such as uPAR and MMP-9.

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Acknowledgements

This work was supported by grants from Murst ex 60% and Ente Cassa di Risparmio di Firenze.

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Authors and Affiliations

  1. Department of Experimental Pathology and Oncology, University of Florence, Florence, Italy

    Chiara Marconi, Francesca Bianchini, Antonella Mannini, Gabriele Mugnai, Salvatore Ruggieri & Lido Calorini

  2. Dipartimento di Patologia e Oncologia Sperimentali, Università degli Studi di Firenze, Viale G.B. Morgagni 50, 50134, Firenze, Italy

    Lido Calorini

Authors
  1. Chiara Marconi
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  2. Francesca Bianchini
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  3. Antonella Mannini
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  4. Gabriele Mugnai
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  5. Salvatore Ruggieri
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  6. Lido Calorini
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Corresponding author

Correspondence to Lido Calorini.

Additional information

This paper was presented in part at the “11th World Congress on Advances in Oncology and 9th International Symposium on Molecular Medicine” 12–14 October 2006, Hersonissos, Crete, Greece.

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Marconi, C., Bianchini, F., Mannini, A. et al. Tumoral and macrophage uPAR and MMP-9 contribute to the invasiveness of B16 murine melanoma cells. Clin Exp Metastasis 25, 225–231 (2008). https://doi.org/10.1007/s10585-007-9136-0

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  • DOI: https://doi.org/10.1007/s10585-007-9136-0

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