Effect of caffeic acid and caffeic acid phenethyl ester on murine osteosarcoma cells: regulation of the NADPH oxidase complex

Detalhes bibliográficos
Autor(a) principal: Pagnan, Ana Lígia
Data de Publicação: 2021
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Biblioteca Digital de Teses e Dissertações da USP
Texto Completo: https://www.teses.usp.br/teses/disponiveis/25/25149/tde-11112021-112933/
Resumo: Osteosarcoma is the most common type of bone cancer among children and adolescents. Metastasis for this cancer happens around 10-25% of the cases, increasing the mortality rate. The search for new therapeutic strategies has increased for phytochemicals due to their potential as antioxidants and anticancer properties. Studies have reported these properties on caffeic acid phenethyl ester (CAPE) and caffeic acid (CA). In this way, the present study aimed to analyze CAPE and CAs anticancer properties on UMR-106 murine osteosarcoma cells after 72 h of treatment. Cell viability was assessed using the MTT and violet crystal reduction assay, with inhibitory concentrations corresponding to 25 and 50% (IC25 and IC50) of 1.3 and 2.7 M for CAPE and 91.0 and 120.0 M for AC, respectively. In addition, a control cell line (MC3T3-E1) was also used for the viabilities assay. The number of apoptotic cells and proliferation rate were quantified by flow cytometry with ANEXIN V-FITC/DRAQ7 and CFSE, respectively and the cell migration behavior was evaluated by the Wound Healing assay. The quantification of reactive oxygen species (ROS) was performed by DCFH-DA fluorescence. NOX-2 and NOX-4 genes were analyzed by RT-qPCR. Data were analyzed by one-way ANOVA. Significant differences between groups were determined by Tukeys post-hoc test at P<0.05. Thus, the present study shows the potential anticancer properties of CAPE and highlights how a simple chemical modification can improve the pharmacological potency of a phytochemical in relation to its precursor CA. Our results showed that CAPE was more efficient and selective in reducing the viability of tumor cells, with significant differences, when compared to the control (P<0.05) and it was 44-fold (IC25) and 70-fold (IC50) more cytotoxic than CA. CAPE also induced apoptosis and decreased ROS generation, in addition to limiting cell migration. In summary, CAPE was more selective for tumor cells, preserving normal ones, suggesting its potential role as an anticancer drug.
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spelling Effect of caffeic acid and caffeic acid phenethyl ester on murine osteosarcoma cells: regulation of the NADPH oxidase complexEfeito do ácido cafeico e do éster fenetil do ácido cafeico em células de osteossarcoma murinos: regulação do complexo NADPH oxidaseÁcido cafeicoCaffeic acidCaffeic ester phenethyl esterCancerCâncerCompostos fenólicosÉster fenetil do ácido cafeicoOsteosarcomaOsteosarcomaPhenolic compoundsOsteosarcoma is the most common type of bone cancer among children and adolescents. Metastasis for this cancer happens around 10-25% of the cases, increasing the mortality rate. The search for new therapeutic strategies has increased for phytochemicals due to their potential as antioxidants and anticancer properties. Studies have reported these properties on caffeic acid phenethyl ester (CAPE) and caffeic acid (CA). In this way, the present study aimed to analyze CAPE and CAs anticancer properties on UMR-106 murine osteosarcoma cells after 72 h of treatment. Cell viability was assessed using the MTT and violet crystal reduction assay, with inhibitory concentrations corresponding to 25 and 50% (IC25 and IC50) of 1.3 and 2.7 M for CAPE and 91.0 and 120.0 M for AC, respectively. In addition, a control cell line (MC3T3-E1) was also used for the viabilities assay. The number of apoptotic cells and proliferation rate were quantified by flow cytometry with ANEXIN V-FITC/DRAQ7 and CFSE, respectively and the cell migration behavior was evaluated by the Wound Healing assay. The quantification of reactive oxygen species (ROS) was performed by DCFH-DA fluorescence. NOX-2 and NOX-4 genes were analyzed by RT-qPCR. Data were analyzed by one-way ANOVA. Significant differences between groups were determined by Tukeys post-hoc test at P<0.05. Thus, the present study shows the potential anticancer properties of CAPE and highlights how a simple chemical modification can improve the pharmacological potency of a phytochemical in relation to its precursor CA. Our results showed that CAPE was more efficient and selective in reducing the viability of tumor cells, with significant differences, when compared to the control (P<0.05) and it was 44-fold (IC25) and 70-fold (IC50) more cytotoxic than CA. CAPE also induced apoptosis and decreased ROS generation, in addition to limiting cell migration. In summary, CAPE was more selective for tumor cells, preserving normal ones, suggesting its potential role as an anticancer drug.O osteossarcoma é o tipo de câncer ósseo mais comum entre crianças e adolescentes. A metástase para esse câncer ocorre em torno de 10 a 25% dos casos, diminuindo a taxa de sobrevivência. A busca por novas estratégias terapêuticas tem aumentado para os fitoquímicos devido ao seu potencial como antioxidantes e propriedades anticâncer. Estudos relataram essas propriedades no éster fenetil do ácido cafeico (CAPE) e no ácido cafeico (AC). Deste modo, o presente trabalho teve como objetivo analisar as propriedades anticâncer de CAPE e AC em células de osteossarcoma murino UMR-106 após 72 horas de tratamento. A viabilidade celular foi avaliada por meio do ensaio de redução do MTT e cristal violeta, sendo as concentrações inibitórias correspondentes a 25 e 50% (IC25 e IC50) de 1.3 e 2.7 M para CAPE e 91.0 e 120.0 M para AC, respectivamente. Adicionalmente, uma linhagem controle (MC3T3-E1) também foi usada para os ensaios de viabilidade. O número de células apoptóticas e a taxa de proliferação foram quantificados por citometria de fluxo com ANEXIN V-FITC/DRAQ7 e CFSE, respectivamente, e o comportamento de migração celular, pelo ensaio de wound healing. A quantificação das espécies reativas de oxigênio (ROS) foi realizada por fluorescência DCFH-DA. Os genes NOX-2 e NOX-4 foram analisados por RT-qPCR. Os dados foram analisados por ANOVA de um fator e diferenças significativas entre os grupos foram determinadas pelo teste post-hoc de Tukey em P<0.05. Sendo assim, o presente trabalho demonstra as potenciais propriedades anticâncer do CAPE e destaca como uma modificação química simples pode melhorar a potência farmacológica de um fitoquímico em relação ao seu precursor AC. Nossos resultados mostraram que o CAPE foi mais eficiente e seletivo na redução da viabilidade das células tumorais, com diferenças significativas, quando comparado ao controle (P<0.05) e foi 44 vezes (IC25) e 70 vezes (IC50) mais citotóxico do que o AC. O CAPE também induziu a apoptose e diminuiu da geração de ROS, além de limitar a migração celular. Em resumo, o CAPE foi mais seletivo para a células tumorais, preservando as normais, sugerindo um papel potencial deste como uma droga anticâncer.Biblioteca Digitais de Teses e Dissertações da USPOliveira, Rodrigo Cardoso deXimenes, Valdecir FariasPagnan, Ana Lígia2021-05-03info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttps://www.teses.usp.br/teses/disponiveis/25/25149/tde-11112021-112933/reponame:Biblioteca Digital de Teses e Dissertações da USPinstname:Universidade de São Paulo (USP)instacron:USPLiberar o conteúdo para acesso público.info:eu-repo/semantics/openAccesseng2024-08-02T12:25:02Zoai:teses.usp.br:tde-11112021-112933Biblioteca Digital de Teses e Dissertaçõeshttp://www.teses.usp.br/PUBhttp://www.teses.usp.br/cgi-bin/mtd2br.plvirginia@if.usp.br|| atendimento@aguia.usp.br||virginia@if.usp.bropendoar:27212024-08-02T12:25:02Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP)false
dc.title.none.fl_str_mv Effect of caffeic acid and caffeic acid phenethyl ester on murine osteosarcoma cells: regulation of the NADPH oxidase complex
Efeito do ácido cafeico e do éster fenetil do ácido cafeico em células de osteossarcoma murinos: regulação do complexo NADPH oxidase
title Effect of caffeic acid and caffeic acid phenethyl ester on murine osteosarcoma cells: regulation of the NADPH oxidase complex
spellingShingle Effect of caffeic acid and caffeic acid phenethyl ester on murine osteosarcoma cells: regulation of the NADPH oxidase complex
Pagnan, Ana Lígia
Ácido cafeico
Caffeic acid
Caffeic ester phenethyl ester
Cancer
Câncer
Compostos fenólicos
Éster fenetil do ácido cafeico
Osteosarcoma
Osteosarcoma
Phenolic compounds
title_short Effect of caffeic acid and caffeic acid phenethyl ester on murine osteosarcoma cells: regulation of the NADPH oxidase complex
title_full Effect of caffeic acid and caffeic acid phenethyl ester on murine osteosarcoma cells: regulation of the NADPH oxidase complex
title_fullStr Effect of caffeic acid and caffeic acid phenethyl ester on murine osteosarcoma cells: regulation of the NADPH oxidase complex
title_full_unstemmed Effect of caffeic acid and caffeic acid phenethyl ester on murine osteosarcoma cells: regulation of the NADPH oxidase complex
title_sort Effect of caffeic acid and caffeic acid phenethyl ester on murine osteosarcoma cells: regulation of the NADPH oxidase complex
author Pagnan, Ana Lígia
author_facet Pagnan, Ana Lígia
author_role author
dc.contributor.none.fl_str_mv Oliveira, Rodrigo Cardoso de
Ximenes, Valdecir Farias
dc.contributor.author.fl_str_mv Pagnan, Ana Lígia
dc.subject.por.fl_str_mv Ácido cafeico
Caffeic acid
Caffeic ester phenethyl ester
Cancer
Câncer
Compostos fenólicos
Éster fenetil do ácido cafeico
Osteosarcoma
Osteosarcoma
Phenolic compounds
topic Ácido cafeico
Caffeic acid
Caffeic ester phenethyl ester
Cancer
Câncer
Compostos fenólicos
Éster fenetil do ácido cafeico
Osteosarcoma
Osteosarcoma
Phenolic compounds
description Osteosarcoma is the most common type of bone cancer among children and adolescents. Metastasis for this cancer happens around 10-25% of the cases, increasing the mortality rate. The search for new therapeutic strategies has increased for phytochemicals due to their potential as antioxidants and anticancer properties. Studies have reported these properties on caffeic acid phenethyl ester (CAPE) and caffeic acid (CA). In this way, the present study aimed to analyze CAPE and CAs anticancer properties on UMR-106 murine osteosarcoma cells after 72 h of treatment. Cell viability was assessed using the MTT and violet crystal reduction assay, with inhibitory concentrations corresponding to 25 and 50% (IC25 and IC50) of 1.3 and 2.7 M for CAPE and 91.0 and 120.0 M for AC, respectively. In addition, a control cell line (MC3T3-E1) was also used for the viabilities assay. The number of apoptotic cells and proliferation rate were quantified by flow cytometry with ANEXIN V-FITC/DRAQ7 and CFSE, respectively and the cell migration behavior was evaluated by the Wound Healing assay. The quantification of reactive oxygen species (ROS) was performed by DCFH-DA fluorescence. NOX-2 and NOX-4 genes were analyzed by RT-qPCR. Data were analyzed by one-way ANOVA. Significant differences between groups were determined by Tukeys post-hoc test at P<0.05. Thus, the present study shows the potential anticancer properties of CAPE and highlights how a simple chemical modification can improve the pharmacological potency of a phytochemical in relation to its precursor CA. Our results showed that CAPE was more efficient and selective in reducing the viability of tumor cells, with significant differences, when compared to the control (P<0.05) and it was 44-fold (IC25) and 70-fold (IC50) more cytotoxic than CA. CAPE also induced apoptosis and decreased ROS generation, in addition to limiting cell migration. In summary, CAPE was more selective for tumor cells, preserving normal ones, suggesting its potential role as an anticancer drug.
publishDate 2021
dc.date.none.fl_str_mv 2021-05-03
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language eng
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dc.rights.driver.fl_str_mv Liberar o conteúdo para acesso público.
info:eu-repo/semantics/openAccess
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eu_rights_str_mv openAccess
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dc.publisher.none.fl_str_mv Biblioteca Digitais de Teses e Dissertações da USP
publisher.none.fl_str_mv Biblioteca Digitais de Teses e Dissertações da USP
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reponame_str Biblioteca Digital de Teses e Dissertações da USP
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repository.name.fl_str_mv Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP)
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