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

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

Autor Toricelli, Mariana Autor UNIFESP Google Scholar
Melo, Fabiana H. M. Autor UNIFESP Google Scholar
Peres, Giovani B. Autor UNIFESP Google Scholar
Silva, Debora C. P. Google Scholar
Jasiulionis, Miriam G. Autor UNIFESP Google Scholar
Instituição Universidade Federal de São Paulo (UNIFESP)
Ludwig Inst Canc Res
Resumo Background: 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.
Palavra-chave Timp1
Beta1-integrin
CD63
PI3-K pathway
Anoikis
Melanoma
Malignant transformation
Idioma Inglês
Financiador Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Número do financiamento FAPESP: 2011/12306-1
FAPESP: 2010/18715-8
CAPES: 2867/10
Data de publicação 2013-03-25
Publicado em Molecular Cancer. London: Biomed Central Ltd, v. 12, 15 p., 2013.
ISSN 1476-4598 (Sherpa/Romeo, fator de impacto)
Publicador Biomed Central Ltd
Extensão 15
Fonte http://dx.doi.org/10.1186/1476-4598-12-22
Direito de acesso Acesso aberto Open Access
Tipo Artigo
Web of Science WOS:000318113300001
Endereço permanente http://repositorio.unifesp.br/handle/11600/36097

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