Introduction: Metastatic potential and rising incidence make cutaneous melanoma a major health concern worldwide. While current therapies have proven effective, they are limited by side effects arising and by the development of drug resistance. For this reason, identifying new therapeutic targets and predictive biomarkers is crucial. Semaphorin 5 A (SEMA5A) is a member of the Semaphorin family, implicated in both cancer biology and neural development. We have previously demonstrated that this protein promotes different aggressive features of melanoma cells. In this study, we conducted a more in-depth investigation into the role of SEMA5A in the development and progression of melanoma. Methods: We performed proteomic and in silico analysis of stable knockdown SEMA5A human melanoma cells to identify the cellular pathways modulated by SEMA5A. The data used are available via ProteomeXchange with the identifier PXD065661. In vitro cell migration, clonogenic ability and cell viability assays under different adhesive substrate conditions were conducted using both human and murine melanoma SEMA5A-depleted clones. Western blotting and immunofluorescence analyses were employed to investigate focal adhesion regulation and lamellipodia formation. A Nocodazole assay was used to evaluate focal adhesion dynamics. A xenograft mouse model of melanoma was used to investigate the role of SEMA5A in tumor formation and tumor growth. Paraffin tumor sections were analyzed using immunohistochemical staining.
Brignone, M., Concetta Cufaro, M., Ercolani, C., Masi, I., Di Ferdinando, F., Palange, A., Valentini, E., Di Martile, M., Del Boccio, P., Del Bufalo, D., Tamagnone, L., Rosanò, L., D’Aguanno, S., Semaphorin 5A modulates focal adhesion pathway and lamellipodia formation in melanoma, <<CELL COMMUNICATION AND SIGNALING>>, 2025; 23 (1): 1-510. [doi:doi: 10.1186/s12964-025-02526-z] [https://hdl.handle.net/10807/329898]
Semaphorin 5A modulates focal adhesion pathway and lamellipodia formation in melanoma
Tamagnone, Luca;
2025
Abstract
Introduction: Metastatic potential and rising incidence make cutaneous melanoma a major health concern worldwide. While current therapies have proven effective, they are limited by side effects arising and by the development of drug resistance. For this reason, identifying new therapeutic targets and predictive biomarkers is crucial. Semaphorin 5 A (SEMA5A) is a member of the Semaphorin family, implicated in both cancer biology and neural development. We have previously demonstrated that this protein promotes different aggressive features of melanoma cells. In this study, we conducted a more in-depth investigation into the role of SEMA5A in the development and progression of melanoma. Methods: We performed proteomic and in silico analysis of stable knockdown SEMA5A human melanoma cells to identify the cellular pathways modulated by SEMA5A. The data used are available via ProteomeXchange with the identifier PXD065661. In vitro cell migration, clonogenic ability and cell viability assays under different adhesive substrate conditions were conducted using both human and murine melanoma SEMA5A-depleted clones. Western blotting and immunofluorescence analyses were employed to investigate focal adhesion regulation and lamellipodia formation. A Nocodazole assay was used to evaluate focal adhesion dynamics. A xenograft mouse model of melanoma was used to investigate the role of SEMA5A in tumor formation and tumor growth. Paraffin tumor sections were analyzed using immunohistochemical staining.
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