Avaliação da atividade antitumoral de compostos n-fenilpiperazínicos em linhagem tumoral K562
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Data
2016-05-20
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Universidade Federal de Goiás
Resumo
Although the efforts employed by scientific community to discover new anticancer therapies
suitable to the increasing cancer incidence and multidrug resistance, its necessary to develop
more selective and target driven drugs. Therefore, in this work we have done a screening with
LQFM030 analogues, which is a Nutlin-1 analogue, aiming to evaluate their cytotoxic
potential. Furthermore, we have evaluated the cytotoxicity, the morphological alterations and
the cell death induction mechanisms of the compound LQFM166 in leukemia cell line K562. In
parallel, we have investigated the security profile of the compound upon 3T3 basal cell line to
estimate its LD50 and the Selectivity Index. The cytotoxicity assays included the tetrazolium
salt (MTT) reduction and the Neutral Red Uptake assays, to assess the cytotoxicity of
LQFM166 in K562 and 3T3 cell lines, respectively. The investigation of cell death induction
mechanisms was carried out using flow cytometry, whereby we have evaluated the cells
biochemistry parameters, including cell cycle progression, phosphatidylserine externalization,
caspases 3/7, 8 and 9 activity, cytochrome c release from mitochondria, p21, p27, Bax, Bcl-2,
cyclin-B1 and NFkB expression, using specific labeling for each assay. Data were analyzed by t
test and the difference between control and treated groups averages was considered
statistically significant when p<0,05. Regarding leukemia cell line K562, the compound
LQFM166 was cytotoxic, showing a dose-time-dependent profile. Morphological alterations
were observed after treatment with the compound at cellular and nuclear levels, which
corroborate with apoptotic cell death. Additionally, treatment with the IC50 for 48 hours has
promoted cellular and molecular changes that characterize this process, including
phosphatidylserine externalization, increase of caspases 3/7, 8 and 9 activity, expression of
pro-apoptotic proteins Bax, p21and p27, as well as diminution of Bcl-2 and cyclin-B1. We
have also observed increase of cytochrome-c release and NFkB expression. Concerning the
security profile, the compound was considered relatively selective, once the IC50 found to
basal cell line (185,3 µM) was the double of the obtained to leukemia cell line regarding the
same time of exposure (56,76 µM). The outcomes allow us to conclude that LQFM166 was
cytotoxic upon leukemia cells K562, promoting morphological and biochemical alterations that
indicate apoptotic cell death induction.
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Palavras-chave
LQFM166 , Trifluormetil , Apoptose , K562 , Morte Celular , LQFM166 , Trifluoromethyl , Apoptosis , K562 , Cell death
Citação
SANTOS, Thaís Rosa Marques dos. Avaliação da atividade antitumoral de compostos n-fenilpiperazínicos em linhagem tumoral K562. 2016. 72 f. Dissertação (Mestrado em Ciências Farmacêuticas) - Universidade Federal de Goiás, Goiânia, 2016.