Background: Invadopodia are actin-based cell-membrane protrusions associated with the extracellular matrix degradation accompanying cancer invasion. The elucidation of the molecular mechanisms leading to invadopodia formation and activity is central for the prevention of tumor spreading and growth. Protein tyrosine kinases such as Src are known to regulate invadopodia assembly, little is however known on the role of protein tyrosine phosphatases in this process. Among these enzymes, we have selected the tyrosine phosphatase Shp1 to investigate its potential role in invadopodia assembly, due to its involvement in cancer development. Methods: Co-immunoprecipitation and immunofluorescence studies were employed to identify novel substrate/s of Shp1AQ controlling invadopodia activity. The phosphorylation level of cortactin, the Shp1 substrate identified in this study, was assessed by immunoprecipitation, in vitro phosphatase and western blot assays. Short interference RNA and a catalytically-dead mutant of Shp1 expressed in A375MM melanoma cells were used to evaluate the role of the specific Shp1-mediated dephosphorylation of cortactin. The anti-invasive proprieties of glycerophosphoinositol, that directly binds and regulates Shp1, were investigated by extracellular matrix degradation assays and in vivo mouse model of metastasis. Results: The data show that Shp1 was recruited to invadopodia and promoted the dephosphorylation of cortactin at tyrosine 421, leading to an attenuated capacity of melanoma cancer cells to degrade the extracellular matrix. Controls included the use of short interference RNA and catalytically-dead mutant that prevented the dephosphorylation of cortactin and hence the decrease the extracellular matrix degradation by melanoma cells. In addition, the phosphoinositide metabolite glycerophosphoinositol facilitated the localization of Shp1 at invadopodia hence promoting cortactin dephosphorylation. This impaired invadopodia function and tumor dissemination both in vitro and in an in vivo model of melanomas. Conclusion: The main finding here reported is that cortactin is a specific substrate of the tyrosine phosphatase Shp1 and that its phosphorylation/dephosphorylation affects invadopodia formation and, as a consequence, the ability of melanoma cells to invade the extracellular matrix. Shp1 can thus be considered as a regulator of melanoma cell invasiveness and a potential target for antimetastatic drugs. [MediaObject not available: see fulltext.]

The phosphatase Shp1 interacts with and dephosphorylates cortactin to inhibit invadopodia function / A. Varone, C. Amoruso, M. Monti, M. Patheja, A. Greco, L. Auletta, A. Zannetti, D. Corda. - In: CELL COMMUNICATION AND SIGNALING. - ISSN 1478-811X. - 19:1(2021), p. 64.64. [10.1186/s12964-021-00747-6]

The phosphatase Shp1 interacts with and dephosphorylates cortactin to inhibit invadopodia function

L. Auletta;
2021

Abstract

Background: Invadopodia are actin-based cell-membrane protrusions associated with the extracellular matrix degradation accompanying cancer invasion. The elucidation of the molecular mechanisms leading to invadopodia formation and activity is central for the prevention of tumor spreading and growth. Protein tyrosine kinases such as Src are known to regulate invadopodia assembly, little is however known on the role of protein tyrosine phosphatases in this process. Among these enzymes, we have selected the tyrosine phosphatase Shp1 to investigate its potential role in invadopodia assembly, due to its involvement in cancer development. Methods: Co-immunoprecipitation and immunofluorescence studies were employed to identify novel substrate/s of Shp1AQ controlling invadopodia activity. The phosphorylation level of cortactin, the Shp1 substrate identified in this study, was assessed by immunoprecipitation, in vitro phosphatase and western blot assays. Short interference RNA and a catalytically-dead mutant of Shp1 expressed in A375MM melanoma cells were used to evaluate the role of the specific Shp1-mediated dephosphorylation of cortactin. The anti-invasive proprieties of glycerophosphoinositol, that directly binds and regulates Shp1, were investigated by extracellular matrix degradation assays and in vivo mouse model of metastasis. Results: The data show that Shp1 was recruited to invadopodia and promoted the dephosphorylation of cortactin at tyrosine 421, leading to an attenuated capacity of melanoma cancer cells to degrade the extracellular matrix. Controls included the use of short interference RNA and catalytically-dead mutant that prevented the dephosphorylation of cortactin and hence the decrease the extracellular matrix degradation by melanoma cells. In addition, the phosphoinositide metabolite glycerophosphoinositol facilitated the localization of Shp1 at invadopodia hence promoting cortactin dephosphorylation. This impaired invadopodia function and tumor dissemination both in vitro and in an in vivo model of melanomas. Conclusion: The main finding here reported is that cortactin is a specific substrate of the tyrosine phosphatase Shp1 and that its phosphorylation/dephosphorylation affects invadopodia formation and, as a consequence, the ability of melanoma cells to invade the extracellular matrix. Shp1 can thus be considered as a regulator of melanoma cell invasiveness and a potential target for antimetastatic drugs. [MediaObject not available: see fulltext.]
No
English
Cancer; Cortactin; Glycerophosphoinositols; Invadopodia; Phosphoinositides; Src homology region 2 domain-containing phosphatase-1 (Shp1); Animals; Cell Line, Tumor; Cortactin; Extracellular Matrix; Humans; Inositol Phosphates; Lung Neoplasms; Melanoma; Mice, Inbred BALB C; Mice, Nude; Models, Biological; Neoplasm Invasiveness; Phosphorylation; Protein Binding; Protein Tyrosine Phosphatase, Non-Receptor Type 6; Pseudopodia; Substrate Specificity
Settore BIO/11 - Biologia Molecolare
Articolo
Sì, ma tipo non specificato
Ricerca di base
Pubblicazione scientifica
2021
BioMed Central Ltd
19
1
64
64
Pubblicato
Periodico con rilevanza internazionale
scopus
wos
pubmed
crossref
datacite
Aderisco
info:eu-repo/semantics/article
The phosphatase Shp1 interacts with and dephosphorylates cortactin to inhibit invadopodia function / A. Varone, C. Amoruso, M. Monti, M. Patheja, A. Greco, L. Auletta, A. Zannetti, D. Corda. - In: CELL COMMUNICATION AND SIGNALING. - ISSN 1478-811X. - 19:1(2021), p. 64.64. [10.1186/s12964-021-00747-6]
open
Prodotti della ricerca::01 - Articolo su periodico
8
262
Article (author)
Periodico con Impact Factor
A. Varone, C. Amoruso, M. Monti, M. Patheja, A. Greco, L. Auletta, A. Zannetti, D. Corda
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/922952
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