Estudo bioanalítico e proteômico de sarcoma de Hippopotumus amphibius

• Ano de defesa: 2014
• Autor(a) principal: Silva, Adriana Rodrigues [UNIFESP]
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de São Paulo
• Palavras-chave em Português: Cancer; Mass spectrometry; Hippopotamus; Proteomics; Sarcoma; Câncer; Espectrometria de massas; Hipopótamo; Proteômica
• Link de acesso: https://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&id_trabalho=2006851; http://repositorio.unifesp.br/handle/11600/46389

This project aims to analyse the protein profile of a Hippopotamus amphibius' sarcoma. Using proteomic analysis on three tissue samples of an animal that passed away due to the advanced stage of the disease, an origin tumor, a heart metastasis and an healthy tissue. Initially, the protein separation was programmed to be performed using bi-dimensional electrophoresis. The small quantity of tissue sample also forced us to use a second methodology, without previous protein separation processes. After the protein samples were handled, they were submitted to the trypsin enzymatic action, followed by purification. The next stage involved the sample analysis using liquid chromatography coupled with mass spectrometry, and data analysis using the Mascot software. The small quantity of tissue samples available made the use of a second methodology necessary, in order to obtain complementary data. Samples from the primary tumor, the heart metastasis, and healthy tissue were submitted to the shot gun, without previous separation of the proteins, going through enzymatic digestion, and the resulting peptides processed with chromatographic separation coupled with mass spectrometry, using the same identification strategy in the first methodology. Though the bi-dimensional gel separation is an expensive and time-costly methodology, it proved to be the more adequate to identify proteins involved in specific biological processes, such as the ones involved in diseases like cancer. The first methodology, that used a protein bi-dimensional gel separation, identified 33 proteins involved in the growth and development of tumors, and the second methodology, that have not used prior separation of the proteins, identified only 8 proteins that are possibly involved in the same cancer related processes. Both methodologies had great value on the development of this study, enabling the work with two strands of proteomics, aiding in the proposed objective, each offering a specific characteristic. The first one was more adequate to the identification of the proteins involved in specific biological processes. The second one, in the ample identification of the proteins expressed in the analyzed tissue. The results can be applied in future studies, that can verify the relation of these proteins with the development and functioning of tumors and the constant improvement of animal conservancy, offering a better understanding of the diseases that affected both animals and humans.