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Previous issue date: 2016-02-17 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / Os exossomos s?o respons?veis pela comunica??o c?lula-c?lula e podem influenciar na progress?o tumoral, met?stase e efic?cia terap?utica. Dentre as c?lulas capazes de secretar exossomos est?o as c?lulas tumorais e as c?lulas imunes. Sabe-se que a presen?a das c?lulas imunes ? importante para erradicar os tumores. No entanto, achados recentes demonstram que a inflama??o pode promover o crescimento tumoral. Os macr?fagos associados a tumores (TAMs) s?o conhecidos por apresentarem diferentes subtipos, M1 e M2, capazes de secretarem exossomos. O presente estudo se prop?s a observar o comportamento dos exossomos derivados dos TAMs, dos subtipos 1 e 2, frente a cultura de c?lulas humanas SCC-25, HSC-3 e SAS derivadas de CE de l?ngua, por meio da an?lise da capacidade de invas?o, prolifera??o e viabilidade das c?lulas tumorais na presen?a dos exossomos. Observou-se que as microves?culas derivadas dos TAMs apresentam positividade para CD63, caracterizando-as como exossomos. Os exossomos dos TAMs do subtipo M2 foram os ?nicos a apresentarem marca??o para TGF-?, quando em compara??o com os exossomos M1, THP1 e das linhagens celulares de CE, sugerindo que os exossomos M2 podem ser respons?veis pela express?o de TGF-? nas c?lulas tumorais, uma vez que s?o internalizados. Nos ensaios de migra??o, observou-se que as c?lulas SCC-25 em presen?a de meio de cultura DMEM F/12, apresentaram maior capacidade de invas?o frente aos exossomos M2 (p?0,001), para concentra??o de 0,1 ?g/ml. Para as c?lulas HSC-3 e SAS, n?o foi observada rela??o estatisticamente significante entre a presen?a de exossomos cultivados juntamente com as c?lulas tumorais e a capacidade de invas?o celular (p>0,05). Quando os exossomos foram colocados no compartimento inferior do transwell, as c?lulas HSC-3 em presen?a dos exossomos M2 (1,0 ?g/ml) apresentaram maior capacidade de invas?o (p?0,001). O teste de viabilidade demonstrou que as c?lulas HSC-3 tornam-se mais vi?veis frente ? presen?a dos exossomos M2 (p?0,001) na concentra??o de 50 ?g/ml. Para as c?lulas SCC-25, o resultado foi o mesmo (p?0,05). A imunofluoresc?ncia demonstrou a internaliza??o dos exossomos nas linhagens celulares estudadas. Os achados sugerem que a presen?a de exossomos M2, frente ?s culturas de c?lulas de CE de l?ngua, pode ser um campo de pesquisa importante para futuros estudos com terapias-alvo. / Squamous cell carcinoma (SCC) of the oral tongue is one of the most common malignant lesions in the oral cavity and is characterized by presenting a locally invasive and aggressive behavior. The exosomes are responsible for cell-cell communication and may influence tumor progression, metastasis and therapeutic efficacy. Among the cells that can secrete exosomes are tumor cells and immune cells. It is known that the presence of immune cells is important to eradicate tumors. However, recent findings suggest that inflammation may promote tumor growth. The tumor associated macrophages (TAMs) are known to have subtypes, M1 and M2, that secrete exosomes. This study?s goal was to observe the behavior of derivatives TAMs exosomes, subtypes 1 and 2, against human cell culture SCC-25, HSC-3 and SAS derived from SCC of oral tongue, through the analysis of invasiveness, proliferation and viability of tumor cells in the presence of exosomes. It was observed that the microvesicles derived from TAMs are positive for CD63, characterizing them as exosomes. The exosomes of the M2 subtype TAMs were the only ones to present TGF-? marking, as compared with M1 exosomes, THP1 and SCC cell lines, suggesting that M2 exosomes may be responsible for TGF-? expression in tumor cells. In the migration tests, it was found that the SCC-25 cells in the presence of culture medium DMEM F / 12 showed higher invasion capacity in the presence of M2 exosomes (p?0,001) to a concentration of 0.1 ?g/ml. For HSC-3 and SAS cells, there was no significant statistical relationship between the presence of exosomes and invasiveness (p> 0.05) for the exosomes derived from TAMs. When the exosomes were placed in the lower compartment of the transwell, the HSC-3 cells in the presence of M2 exosomes (1.0 ?g/ml) had higher invasiveness (p?0,001). The viability test showed that HSC-3 cells became more viable in the presence of M2 exosomes (p?0.001) at a concentration of 50 ?g/ml. For SCC-25 cells, the result was the same (p?0.05). Immunofluorescence showed the internalization of exosomes in the cell lines studied. These findings suggest that the presence of M2 exosomes in the SCC of the oral tongue cell cultures may be an important research field for future studies of targeted therapies.
| Identifer | oai:union.ndltd.org:IBICT/oai:repositorio.ufrn.br:123456789/21436 |
| Date | 17 February 2016 |
| Creators | Demeda, Clarissa Favero |
| Contributors | 01608789420, http://lattes.cnpq.br/7040457616034306, Barboza, Carlos Augusto Galv?o, 67269621420, http://lattes.cnpq.br/5004397230198722, Korvala, Johanna Katariina, 00000000000, Souza, Lelia Batista de, 08588724472, http://lattes.cnpq.br/6671914892609743, Colleta, Ricardo Della, 13957746841, Pinto, Le?o Pereira |
| Publisher | PROGRAMA DE P?S-GRADUA??O EM PATOLOGIA ORAL, UFRN, Brasil |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/doctoralThesis |
| Source | reponame:Repositório Institucional da UFRN, instname:Universidade Federal do Rio Grande do Norte, instacron:UFRN |
| Rights | info:eu-repo/semantics/openAccess |
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