Banca de DEFESA: JONAS BISPO PESSOA

Uma banca de DEFESA de MESTRADO foi cadastrada pelo programa.
DISCENTE : JONAS BISPO PESSOA
DATA : 06/12/2016
HORA: 09:30
LOCAL: Sala de Reuniões DMOR
TÍTULO:

Evaluation of the effects of the synergistic action of the antihypertensive carvedilol and gold nanoparticle on human hepatocellular carcinoma cells.

PALAVRAS-CHAVES:

Liver cancer, Apoptosis, Gold nanoparticle, Carvedilol.

PÁGINAS: 85
GRANDE ÁREA: Ciências Biológicas
ÁREA: Farmacologia
SUBÁREA: Farmacologia Bioquímica e Molecular
RESUMO:

Liver cancer is considered one of the most prevalent in the world and does not present promising treatments, necessitating alternative forms of fighting the tumor. Gold nanoparticles have emerged as an important modality of treatment for various diseases due to the characteristics of their nanoconjugates. In addition to this, the antihypertensive carvedilol has been shown in the recent literature to be antitumor potential. Therefore, the objective of this study was to evaluate the antitumor action, isolated and synergistic, of the gold nanoparticle and the antihypertensive carvedilol on tumor cells (HepG2) and human normal (HEK-293). The viability test was carried out by exclusion of tripan blue at doses of NPO (1 μg / ml, 3 μg / ml, 6.25 μg / ml, 12.5 μg / ml, 25 μg / ml and 50 μg / Ml) and carvedilol (1.5 μM, 3 μM, 6.25 μM, 12.5 μM, 50 μM, 100 μM, 200 μM and 300 μM) were used to select those that would cause low inhibition of growth of HepG2 cells. The selected doses of NPO and carvedilol were used, isolated and in synergism, for analysis of cell death by flow cytometry by the labeling of Anexins V-FITC and propidium iodide (PI). Subsequently, tumor cells were analyzed for expression of caspase-3, Bcl-2, caspase-8 and MAPK / ERK proteins by immunofluorescence microscopy. Then, mRNA levels of FADD, Apaf-1, survivin, MDR-1, EGFR, Akt and mTOR were measured by relating them to resistance and cell death. The evaluation of intracellular NPO targets, isolated and in synergism, was performed by transmission electron microscopy (TEM). Observing the synergy, the cells were submitted to gold nanoparticle and 24 hours after the treatment, treated with carvedilol. The best doses of the cell viability test with NPO (3 μg / ml and 6.25 μg / ml) and carvedilol (1.5 μM and 3 μM) showed, by flow cytometry, pro-apoptotic activity on cells (HepG2) with statically significant results for synergism (P <0.001). Relative to normal human cells (HEK-293), these same doses did not prove to be statistically significant apoptosis promoters at both times. There was a reduction in total apoptosis (P <0.05) for the highest dose of synergism. There were strong markers for caspase-3 and caspase-8, in the groups treated with NPO (6.25 μg / m) and carvedilol (3 μM), alone and in synergism, and lack of labeling for Bcl-2 and MAPK / ERK for The same groups. In addition, mRNA expression of anti-apoptotic proteins (EGFR Akt, mTOR) and resistance (MDR-1) was shown to be overregulated, while the gene expression of proapoptotic proteins (FADD) was overregulated . MET demonstrated the internalization of NPO alone, in the vicinity of the plasma membrane and, in synergism, in the vicinity of the nucleus. With this, it can be concluded that the synergistic action of NPO and carvedilol shows proapoptotic action on tumor cells and protection of normal cells.

MEMBROS DA BANCA:
Externo ao Programa - 2374605 - AURIGENA ANTUNES DE ARAUJO
Externo à Instituição - JEYMESSON RAPHAEL CARDOSO VIEIRA - UFPE
Presidente - 2329140 - RAIMUNDO FERNANDES DE ARAUJO JUNIOR