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Log inINTRODUCTION AND OBJECTIVES: Glioblastoma (GBM) is the most invasive malignant tumors in the Central Nervous System. Marine bioprospecting is a relevant anti-tumoral strategy in natural products field. Ascidian compounds have already found applications in cancer treatment, neurodegenerative and other diseases. The aim of this study is to evaluate cytotoxic and cytostatic effect of the ascidian Didemnum sp. extract and isolated/combined compounds in glioblastoma cells.
MATERIAL AND METHODS: We used the dynamic maceration process with 1:5 proportion methanol as solvent. After filtration, the solvent was evaporated at 50oC in a water bath, obtaining Didemnum sp. pull extract sample. Cytosolic acid phosphatase activity was used for cell viability assay by spectrophotometry and cell cycle was observed by flow cytometry. GBM cell lines T98G and U251, and a healthy cell line of human fibroblast BJ-5ta (IHF) were tested in a monolayer model, using increasing concentrations of the Didemnum sp. extract for 72h. Cell morphology was observed by contrast phase and DAPI staining.
RESULTS AND CONCLUSION: Didemnum sp. crude extract demonstrated significant anti-tumoral activity for T98G and U251 monolayers, with IC50 of 120.6 µg/mL and 63.32 µg/mL, respectively. Also, there was a relevant reduction of total DAPI+ cells percentual in both cell lines, and no cytotoxicity in IHF cells. However, no alteration in cell cycle phases was demonstrated. Such effectiveness demonstrates the anti-tumor potential of the Didemnum sp. extract. We expect to isolate the alkaloids and fatty acid identified by LC/MS/MS chromatography to test compound association strategies comparing to the crude extract in healthy and tumoral cells of the CNS in vitro, by spheroid model, and in vivo, by orthotopic model in C57BL6 mice. We suggest marine bioproducts candidates for GBM treatment as new and selective strategies in the future.
Fabio Hecht Castro Medeiros
Boa tarde, Bianca. Parabéns pelo trabalho.
Dúvida 1) você mencionou que o extrato bruto não teve nenhuma toxicidade em fibroblastos. Que dados são esses? Por que eles não foram mostrados? Eles já existem ou você ainda pretende fazer? Isso não ficou claro pra mim
Dúvida 2) Na metodologia você diz que "o ciclo celular foi avaliado por citometria de fluxo". Também menciona a coloração com DAPI. Qual a intenção da coloração com DAPI? E mais importante, onde estão esses dados?
Dúvida 3) Você menciona como objetivo futuro fazer experimentos in vivo com compostos isolados. Imagino que depois de caracterizar quais compostos tem atividade citotóxica. Considerando que isso já tenha sido feito, como vc pretende fazer os experimentos in vivo? Já sabe qual modelo irá usar?
Elaine da Conceição Petronilho
Nas suas perspectivas vocês pretendem extrair e analisar alcalóides e ácidos graxos da Didemnun sp. Os alcalóides são conhecidos na literatura por suas atividades biológicas e químicas. Qual atividade vocês esperam ver em relação aos ácidos graxos? Por que analisar este tipo de composto frente ao GBM?
Elaine da Conceição Petronilho
Qual o controle utilizado? Usaram algum composto padrão que possua um IC50 considerado efetivo para essas células testadas ou só compararam com a literatura? Caso tenha sido por comparação, qual o resultado obtiveram?
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