Detalhes bibliográficos
| Ano de defesa: |
2018 |
| Autor(a) principal: |
Oliveira, Jéssica Gisleine de
 |
| Orientador(a): |
Zuccari, Debora Aparecida Pires de Campos
|
| Banca de defesa: |
Antonucci, Gilmara Ausech
,
Pavarino, Érika Cristina
|
| Tipo de documento: |
Dissertação
|
| Tipo de acesso: |
Acesso aberto |
| Idioma: |
por |
| Instituição de defesa: |
Faculdade de Medicina de São José do Rio Preto
|
| Programa de Pós-Graduação: |
Programa de Pós-Graduação em Ciências da Saúde
|
| Departamento: |
Faculdade 1::Departamento 1
|
| País: |
Brasil
|
| Palavras-chave em Português: |
|
| Palavras-chave em Inglês: |
|
| Área do conhecimento CNPq: |
|
| Link de acesso: |
http://bdtd.famerp.br/handle/tede/538
|
Resumo: |
Breast cancer presents high incidence and death rates worldwide. Metastasis is one of the main causes of high mortality rates by this neoplasm. MicroRNAs (miRNAs) are small non-coding RNA molecules with nearly 18-22 nucleotides, involved in gene expression regulation. Studies demonstrate the role this molecules in tumor progression, including breast cancer. Melatonin, a hormone secreted mainly in pineal gland, has been presenting several effects in cancer regulation, acting on the modulation of proteins and miRNAs. Aim: The aim of the present study was to investigate the action of melatonin on miRNA-10a-5p modulation in triple-negative breast cancer cells (TNBC), and associating with tumor progression. Materials and Methods: The MTT (3- (4,5-Dimethylthiazol-2-yl) -2,5-Diphenyltetrazolium Bromide) assay was performed to verify cell viability of MDA-MB-468 cells at different concentrations of melatonin for 24 hours. The differential expression of 84 miRNAs associated with breast cancer was verified by PCR-Array. The miR-10a was selected for its involvement with tumor process and its gene expression and its target HOXD10 were verified by real-time PCR in breast cancer cell lines. The following groups were considered: cells treated or not with melatonin; transfected with inhibition of miR-10a or negative control of reaction. The invasion and migration assay using matrigel inserts was performed and protein expression of epithelial-mesenchymal transition (EMT), Ecadherin, claudin 7 and vimentin markers were investigated, as PIK3CA proliferation marker, by Western blotting. Results: Our results showed that 1 mM melatonin significantly decreased cell viability. The results of PCR Array showed thirteen miRNAs modulated by melatonin (six overexpressed and seven inhibited). The gene expression showed a decrease in relative amount of miR-10a and its target HOXD10 after melatonin treatmet. The matrigel assays showed that melatonin and inhibition of miR-10a decreased the cell invasion and migration. According to the protein analysis, melatonin was able to reduce the expression of vimentin and claudin 7 proteins and increase E-cadherin expression. On the other hand, inhibition of miR-10a reduced the vimentin protein and did not modulate claudin 7 and E-cadherin. Conclusion: In this study was verified the ability of melatonin modulating miRNAs expression by decreasing miR-10a, affecting tumor invasion and migration, and proteins involved with EMT. In this way, we support the idea of the potential role of melatonin in the regulation of metastasis. |
