Timp1 interacts with beta-1 integrin and CD63 along melanoma genesis and confers anoikis resistance by activating PI3-K signaling pathway independently of Akt phosphorylation

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dc.contributor.authorToricelli, Mariana [UNIFESP]
dc.contributor.authorMelo, Fabiana H. M. [UNIFESP]
dc.contributor.authorPeres, Giovani B. [UNIFESP]
dc.contributor.authorSilva, Debora C. P.
dc.contributor.authorJasiulionis, Miriam G. [UNIFESP]
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.contributor.institutionLudwig Inst Canc Res
dc.date.accessioned2016-01-24T14:31:26Z
dc.date.available2016-01-24T14:31:26Z
dc.date.issued2013-03-25
dc.description.abstractBackground: Anoikis resistance is one of the abilities acquired along tumor progression. This characteristic is associated with metastasis development, since tumorigenic cells must survive independently of cell-matrix interactions in this process. in our laboratory, it was developed a murine melanocyte malignant transformation model associated with a sustained stressful condition. After subjecting melan-a melanocytes to 1, 2, 3 and 4 cycles of anchorage impediment, anoikis resistant cells were established and named 1C, 2C, 3C and 4C, respectively. These cells showed altered morphology and PMA independent cell growth, but were not tumorigenic, corresponding to pre-malignant cells. After limiting dilution of 4C pre-malignant cells, melanoma cell lines with different characteristics were obtained. Previous data from our group showed that increased Timp1 expression correlated with anoikis-resistant phenotype. Timp1 was shown to confer anchorage-independent growth capability to melan-a melanocytes and render melanoma cells more aggressive when injected into mice. However, the mechanisms involved in anoikis regulation by Timp1 in tumorigenic cells are not clear yet.Methods: the beta 1-integrin and Timp1 expression were evaluated by Western blotting and CD63 protein expression by flow cytometry using specific antibodies. To analyze the interaction among Timp1, CD63 and beta 1-integrin, immunoprecipitation assays were performed, anoikis resistance capability was evaluated in the presence or not of the PI3-K inhibitors, Wortmannin and LY294002. Relative expression of TIMP1 and CD63 in human metastatic melanoma cells was analyzed by real time PCR.Results: Differential association among Timp1, CD63 and beta 1-integrins was observed in melan-a melanocytes, 4C pre-malignant melanocytes and 4C11- and 4C11+ melanoma cells. Timp1 present in conditioned medium of melanoma cells rendered melan-a melanocytes anoikis-resistant through PI3-K signaling pathway independently of Akt activation. in human melanoma cell lines, in which TIMP1 and beta-1 integrin were also found to be interacting, TIMP1 and CD63 levels together was shown to correlate significantly with colony formation capacity.Conclusions: Our results show that Timp1 is assembled in a supramolecular complex containing CD63 and beta 1-integrins along melanoma genesis and confers anoikis resistance by activating PI3-K signaling pathway, independently of Akt phosphorylation. in addition, our data point TIMP1, mainly together with CD63, as a potential biomarker of melanoma.en
dc.description.affiliationUniversidade Federal de São Paulo, Dept Pharmacol, São Paulo, Brazil
dc.description.affiliationUniversidade Federal de São Paulo, Microbiol Immunol & Parasitol Dept, São Paulo, Brazil
dc.description.affiliationUniversidade Federal de São Paulo, Dept Biochem, São Paulo, Brazil
dc.description.affiliationLudwig Inst Canc Res, São Paulo, Brazil
dc.description.affiliationUnifespUniversidade Federal de São Paulo, Dept Pharmacol, São Paulo, Brazil
dc.description.affiliationUnifespUniversidade Federal de São Paulo, Microbiol Immunol & Parasitol Dept, São Paulo, Brazil
dc.description.affiliationUnifespUniversidade Federal de São Paulo, Dept Biochem, São Paulo, Brazil
dc.description.sourceWeb of Science
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
dc.description.sponsorshipIDFAPESP: 2011/12306-1
dc.description.sponsorshipIDFAPESP: 2010/18715-8
dc.description.sponsorshipIDCAPES: 2867/10
dc.format.extent15
dc.identifierhttp://dx.doi.org/10.1186/1476-4598-12-22
dc.identifier.citationMolecular Cancer. London: Biomed Central Ltd, v. 12, 15 p., 2013.
dc.identifier.doi10.1186/1476-4598-12-22
dc.identifier.fileWOS000318113300001.pdf
dc.identifier.issn1476-4598
dc.identifier.urihttp://repositorio.unifesp.br/handle/11600/36097
dc.identifier.wosWOS:000318113300001
dc.language.isoeng
dc.publisherBiomed Central Ltd
dc.relation.ispartofMolecular Cancer
dc.rightsinfo:eu-repo/semantics/openAccess
dc.subjectTimp1en
dc.subjectBeta1-integrinen
dc.subjectCD63en
dc.subjectPI3-K pathwayen
dc.subjectAnoikisen
dc.subjectMelanomaen
dc.subjectMalignant transformationen
dc.titleTimp1 interacts with beta-1 integrin and CD63 along melanoma genesis and confers anoikis resistance by activating PI3-K signaling pathway independently of Akt phosphorylationen
dc.typeinfo:eu-repo/semantics/article
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