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Master's Dissertation
DOI
https://doi.org/10.11606/D.25.2020.tde-22102021-132403
Document
Author
Full name
Adriano de Souza Pessôa
Institute/School/College
Knowledge Area
Date of Defense
Published
Bauru, 2020
Supervisor
Committee
Oliveira, Rodrigo Cardoso de (President)
Damante, Carla Andreotti
Garlet, Ana Paula Favaro Trombone
Zambuzzi, Willian Fernando
Title in Portuguese
Efeito do Vanilato de Metila e Divanilato de Metila em células de câncer de mama: Regulação do complexo NADPH oxidase
Keywords in Portuguese
Divanilato de metila
Pró-oxidante.
Vanilato de metila
Abstract in Portuguese
O câncer de mama é o câncer mais comum entre as mulheres. A busca por novas estratégias de tratamento menos invasivas e danosas ao organismo são bastante estudadas. Neste contexto, os fitoquímicos e seus derivados são conhecidos por exibir uma ampla gama de atividades bioquímicas e farmacológicas benéficas. Assim, este estudo avaliou os efeitos citotóxicos e anticâncer do vanilato de metila (MV) e divanilato de metila (DMV) nas linhagens celulares de câncer MCF-7 e MDA-MB-231, dependente de estrogênio e triplo negativo, respectivamente. Primeiramente, as concentrações de 250 a 4000 M de MV e 25 a 400 M de DMV foram testados pelo ensaio MTT nas linhagens celulares HB4a (linhagem celular de mama normal - controle), MCF-7 e MDA-MB-231. Em seguida, as células foram tratadas com 780 M de VM e 100 M de DMV após 72 h para avaliar a eficiência anticâncer. As células foram coradas com cristal violeta para avaliar a viabilidade. A morfologia celular foi verificada pelas colorações de Hematoxilina/Eosina e DAPI/Faloidina. O número de células apoptóticas e a proliferação foram analisados por citometria de fluxo. A quantificação da produção de espécies reativas de oxigênio (EROs) foi realizada usando DCFH-DA por fluorescência. A expressão dos genes NADPH oxidases (NOXs) foi avaliada por RT-qPCR. Os dados foram analisados por one-way ANOVA. Diferenças significativas entre os grupos foram determinadas pelo teste post-hoc de Dunnett com p <0,05. Os resultados demonstram maior diminuição de viabilidade celular pelo DMV, mostrando maior efeito apoptótico e inibição de proliferação celular em comparação ao VM, em ambas as linhas celulares. Os compostos mostraram ação pró-oxidante e sugerem não regular o complexo NADPH oxidase. O dímero mostrou maior efetividade e efeito pró-oxidante na linhagem MCF-7 que MDA-MB-231 usando este modelo experimental.
Title in English
Effect of Methyl Vanillate and Methyl Divanillate on breast cancer cells: regulation of the NADPH oxidase complex
Keywords in English
Methyl divanillate
Methyl vanillate
Pro-oxidant
Abstract in English
Breast cancer is the most common cancer among women. The search for new strategies for less invasive treatments and harmful to the body are widely studied. In this context, phytochemicals and their derivatives are known to exhibit a wide range of beneficial biochemical and pharmacological activities. This study evaluated the cytotoxic and anticancer effects of Methyl Vanillate (MV) and Methyl Divanillate (DMV) compounds on MCF-7 and MDA-MB-231 cancer cell lines, estrogen dependent and triple negative, respectively. Firstly, the following concentrations 250 to 4000 M of MV; 25 to 400 M of DMV on HB4a (control line), MCF-7 and MDA-MB-231 cell lines were tested by MTT assay. Then, the cells were treated with 780 M of the VM and 100 M of the DMV after 72h to evaluate the anticancer efficiency . The cells were stained using Crystal Violet to evaluate of viability. The morphology of cell was verified by Hematoxylin/Eosin and DAPI/Phalloidin stains. The number of apoptotic cells were quantified by flow cytometry. The proliferation of cells was analyzed by flow cytometry. The ROS production was performed using DCFH-da by fluorescence. NOX gene expression was evaluated by RT-qPCR. Data were analyzed by one-way ANOVA. Significant differences between groups were determined by Dunnett's post-hoc test at p < 0.05. DMV decreased the viability cell, showing higher apoptotic effect and inhibition of proliferation cell compared to VM, on both cell lines. The compounds have shown pro-oxidant action and did not regulate the NADPH oxidase complex. The dimer showed more effectivity and pro-oxidant effect on cell line MCF-7 then MDA-MB-231 using this experimental model.
 
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Release Date
2023-10-22
Publishing Date
2021-10-22
 
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