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Doctoral Thesis
DOI
10.11606/T.42.2015.tde-24082015-185603
Document
Author
Full name
Marcel Shiniti Urabayashi
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2015
Supervisor
Committee
Santelli, Glaucia Maria Machado (President)
Custódio, Márcio Reis
Ferro, Emer Suavinho
Ortis, Fernanda
Silva Junior, Pedro Ismael da
Title in Portuguese
Prospecção de moléculas bioativas em esponjas marinhas da espécie Amphimedon viridis: estudos celulares e moleculares.
Keywords in Portuguese
Amphimedon viridis
Citotoxicidade
Esponja marinha
Moléculas bioativas
RNA
Transcriptoma
Abstract in Portuguese
A análise química do extrato bruto de esponja marinha é um método clássico de descoberta de novas drogas. Metabólitos secundários e peptídeos são geralmente os resultados neste tipo de pesquisa, utilizando diferentes e consecutivos protocolos de purificação. Neste estudo de uma amostra de extrato aquoso esponja separada numa coluna Sephadex®-G25 foi purificado por HPLC, numa tentativa de isolar uma única molécula. Com uma base de dados de RNA e a análise in silico espera-se que o RNAm pode ser relacionado com a molécula precursora do composto bioativo ou mesmo o seu gene. A dissociação de células a partir da matriz da esponja é um método importante para a extração de RNA optimizado em quantidade e qualidade. Frações extrato bruto do organismo e dados de RNAm a partir de células isoladas da esponja podem ser comparados por uma abordagem de comparação dos dados. Ensaios de MTS foram utilizados para analisar a bioatividade e microscopia confocal foi a principal ferramenta para examinar os danos em células tumorais (T47D).
Title in English
Prospection of bioative molecules in Amphimedon viridis sea sponges species: cell and molecular studies.
Keywords in English
Amphimedon viridis
Bioactive molecules
Cytotoxicity
RNA
Sea sponge
Transcriptome
Abstract in English
The chemical analysis of sea sponge crude extract is a classic method in drug discovery. Secondary metabolites and peptides are usually the results in this type of research using different and consecutives purification protocols. In this study a sample of sponge aqueous extract separated in a sephadex-G25 column was purified by HPLC in an attempt to isolate a single molecule. With a RNA databank and in silico analysis is expected that the mRNA may be related with the precursor molecule of the bioactive compound or even its gene. The dissociation of cells from the sponge matrix was an important method to the optimized RNA extraction in quantity and quality. Crude extract fractions of the organism and mRNA data from sponge-isolated cells were obtained to a next step cross-data approach. MTS assays were used to analyze the bioactivity and confocal microscopy was the main tool to scan the damage in tumor cells (T47D).
 
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Publishing Date
2015-08-28
 
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