Extrato de chá verde (Camellia sinensis): um potencial agente antitumoral em células de câncer de mama

Detalhes bibliográficos
Autor(a) principal: Santos, Ronimara Aparecida
Data de Publicação: 2018
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações da UERJ
Texto Completo: http://www.bdtd.uerj.br/handle/1/7244
Resumo: Green tea (GT), a beverage derived from Camellia sinensis, has been extensively studied for its antioxidant property, which is attributed to catechins. These phenolic compounds have been shown to inhibit carcinogenesis, but its mechanisms are still poorly understood. Additionally, most studies perform assays using isolated polyphenols from GT. We aimed to obtain a catechin-rich green tea extract (GTE) and to evaluate its antitumor potential on human breast cancer. Tea infusion was prepared on a ratio of 1 g: 40 mL of boiling distilled water using a combination of temperature and time (70, 75, 80, 85, 90, 95 and 100 ºC/5, 10, 15 min). Infusions were evaluated by polyphenols content (Folin Ciocalteu s reagent) and antioxidant potential (FRAP). Breast cancer cells (MDA-MB-231 and MCF-7) and non-tumoral cells (MCF-10A), were exposed to different concentrations of GTE (31.5 μg/mL to 1.0 mg/mL) during 24 h-48 h, and cell viability was assessed by Alamar Blue® and Trypan Blue assays. Viability was also acessed in the presence of p53 inhibitor, pifithrin-α. Immunocytochemistry and Western blotting analysis were performed to access the expression of p53. Our results demonstrated no influence of extraction condition in the total content of polyphenols and antioxidant potential of GT, and a new extract was obtained at 80 ºC/5 min, then freeze dried to be used in cell culture. After exposure for 24h, GTE already promoted a cytotoxic effect, reducing viability of both breast cancer cell lines (IC50 MDA-MB-231 = 138,8 μg/mL; IC50 MCF-7 = 324,5 μg/mL) without cytotoxic effect on normal cells (IC50 MCF-10A = 10097 μg/mL). In addition, GTE promotes an increase of p53 levels on treated MCF-7 cells, which express the wild-type form of the protein. Interestingly, an opposite phenomenon was evidenced in MDA-MB-231 cells, that express the mutated form of p53, in which protein levels seem to be lower in the presence of the extract. Our data suggests that GTE has anticarcinogenic potential on breast cancer and it is possible that this capacity might be related with the modulation of p53
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spelling Costa, Danielly Cristiny Ferraz dahttp://lattes.cnpq.br/9617293991265023Daleprane, Júlio Beltramehttp://lattes.cnpq.br/6027048461430935Rangel, Luciana Pereirahttp://lattes.cnpq.br/6987913099524130Gimba, Etel Rodrigues Pereirahttp://lattes.cnpq.br/8976892755529994http://lattes.cnpq.br/5223038433573741Santos, Ronimara Aparecida2021-01-05T16:42:19Z2018-12-102018-07-19SANTOS, Ronimara Aparecida. Extrato de chá verde (Camellia sinensis): um potencial agente antitumoral em células de câncer de mama. 2018. 22 f. Dissertação (Mestrado em Alimentação, Nutrição e Saúde) - Universidade do Estado do Rio de Janeiro, Rio de Janeiro, 2018.http://www.bdtd.uerj.br/handle/1/7244Green tea (GT), a beverage derived from Camellia sinensis, has been extensively studied for its antioxidant property, which is attributed to catechins. These phenolic compounds have been shown to inhibit carcinogenesis, but its mechanisms are still poorly understood. Additionally, most studies perform assays using isolated polyphenols from GT. We aimed to obtain a catechin-rich green tea extract (GTE) and to evaluate its antitumor potential on human breast cancer. Tea infusion was prepared on a ratio of 1 g: 40 mL of boiling distilled water using a combination of temperature and time (70, 75, 80, 85, 90, 95 and 100 ºC/5, 10, 15 min). Infusions were evaluated by polyphenols content (Folin Ciocalteu s reagent) and antioxidant potential (FRAP). Breast cancer cells (MDA-MB-231 and MCF-7) and non-tumoral cells (MCF-10A), were exposed to different concentrations of GTE (31.5 μg/mL to 1.0 mg/mL) during 24 h-48 h, and cell viability was assessed by Alamar Blue® and Trypan Blue assays. Viability was also acessed in the presence of p53 inhibitor, pifithrin-α. Immunocytochemistry and Western blotting analysis were performed to access the expression of p53. Our results demonstrated no influence of extraction condition in the total content of polyphenols and antioxidant potential of GT, and a new extract was obtained at 80 ºC/5 min, then freeze dried to be used in cell culture. After exposure for 24h, GTE already promoted a cytotoxic effect, reducing viability of both breast cancer cell lines (IC50 MDA-MB-231 = 138,8 μg/mL; IC50 MCF-7 = 324,5 μg/mL) without cytotoxic effect on normal cells (IC50 MCF-10A = 10097 μg/mL). In addition, GTE promotes an increase of p53 levels on treated MCF-7 cells, which express the wild-type form of the protein. Interestingly, an opposite phenomenon was evidenced in MDA-MB-231 cells, that express the mutated form of p53, in which protein levels seem to be lower in the presence of the extract. Our data suggests that GTE has anticarcinogenic potential on breast cancer and it is possible that this capacity might be related with the modulation of p53O chá verde (CV) (Camellia sinensis) tem sido amplamente estudado por sua propriedade antioxidante, atribuída às suas catequinas. Estes compostos fenólicos demonstraram potencial de inibir a carcinogênese, porém os mecanismos envolvidos não estão completamente esclarecidos. Nosso objetivo foi obter um extrato de chá verde (ECV) rico em compostos fenólicos e avaliar seu potencial antitumoral no câncer de mama humano. A infusão de chá foi preparada na proporção de 1 g:40 mL de água utilizando combinações de tempo/temperatura (70-100 ºC/5-15 min). As infusões foram avaliadas pelo conteúdo de polifenóis (Folin-Ciocalteu) e pelo potencial antioxidante (FRAP). Células de câncer de mama (MDA-MB-231 e MCF-7) e células não tumorais mamárias (MCF-10A) foram expostas a diferentes concentrações de ECV (31,5 μg/mL a 1,0 mg/mL) durante 24 h-48 h, e a viabilidade celular foi avaliada (Alamar Blue® e Azul de Tripan). Ensaios de viabilidade celular na presença de pifitrina-α, um inibidor da proteína supressora de tumores p53, também foram conduzidos. Análises de imunocitoquímica e Western bllotting foram realizadas para determinar a expressão de p53. Nossos resultados demonstraram que não houve influência da condição de extração sobre o conteúdo total de polifenóis e potencial antioxidante do CV, sendo um novo extrato obtido a 80 ºC/5 min e liofilizado para aplicação em cultura de células. Após exposição por 24 h, o ECV foi eficaz para reduzir a viabilidade de ambas as linhagens tumorais, sendo mais efetivo sobre a linhagem MDA-MB-231 (IC50 MDA-MB-231 = 138,8 μg/mL; IC50 MCF-7 = 324,5 μg/mL), sem produzir efeito citotóxico em células não tumorais (IC50 MCF-10A = 10097 μg/mL). Além disso, o ECV aumentou os níveis de p53 em células MCF-7, que expressam a forma selvagem desta proteína. Um efeito oposto foi observado na linhagem MDA-MB-231, alterada para p53, na qual seus níveis parecem reduzidos. Todos esses achados sugerem que o ECV apresenta potencial anticarcinogênico sobre células tumorais de mama e, possivelmente, esse fenômeno está relacionado à modulação da proteína p53Submitted by Boris Flegr (boris@uerj.br) on 2021-01-05T16:42:19Z No. of bitstreams: 1 Dissert_Ronimara Aparecida Santos.pdf: 563604 bytes, checksum: 9f4abd378c55a493322cec67919a5f53 (MD5)Made available in DSpace on 2021-01-05T16:42:19Z (GMT). No. of bitstreams: 1 Dissert_Ronimara Aparecida Santos.pdf: 563604 bytes, checksum: 9f4abd378c55a493322cec67919a5f53 (MD5) Previous issue date: 2018-07-19Conselho Nacional de Desenvolvimento Científico e Tecnológicoapplication/pdfporUniversidade do Estado do Rio de JaneiroPrograma de Pós-Graduação em Alimentação, Nutrição e SaúdeUERJBRCentro Biomédico::Instituto de NutriçãoGreen teaPoliphenolsCâncerPolifenóisp53NutriçãoChá verdeCâncerCNPQ::CIENCIAS DA SAUDE::NUTRICAO::DESNUTRICAO E DESENVOLVIMENTO FISIOLOGICOExtrato de chá verde (Camellia sinensis): um potencial agente antitumoral em células de câncer de mamaGreen tea extrat (Camellia sinensis): a potential antitumoral agent on breast cancer cellsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações da UERJinstname:Universidade do Estado do Rio de Janeiro (UERJ)instacron:UERJORIGINALDissert_Ronimara Aparecida Santos.pdfapplication/pdf563604http://www.bdtd.uerj.br/bitstream/1/7244/1/Dissert_Ronimara+Aparecida+Santos.pdf9f4abd378c55a493322cec67919a5f53MD511/72442024-02-23 16:32:22.443oai:www.bdtd.uerj.br:1/7244Biblioteca Digital de Teses e Dissertaçõeshttp://www.bdtd.uerj.br/PUBhttps://www.bdtd.uerj.br:8443/oai/requestbdtd.suporte@uerj.bropendoar:29032024-02-23T19:32:22Biblioteca Digital de Teses e Dissertações da UERJ - Universidade do Estado do Rio de Janeiro (UERJ)false
dc.title.por.fl_str_mv Extrato de chá verde (Camellia sinensis): um potencial agente antitumoral em células de câncer de mama
dc.title.alternative.eng.fl_str_mv Green tea extrat (Camellia sinensis): a potential antitumoral agent on breast cancer cells
title Extrato de chá verde (Camellia sinensis): um potencial agente antitumoral em células de câncer de mama
spellingShingle Extrato de chá verde (Camellia sinensis): um potencial agente antitumoral em células de câncer de mama
Santos, Ronimara Aparecida
Green tea
Poliphenols
Câncer
Polifenóis
p53
Nutrição
Chá verde
Câncer
CNPQ::CIENCIAS DA SAUDE::NUTRICAO::DESNUTRICAO E DESENVOLVIMENTO FISIOLOGICO
title_short Extrato de chá verde (Camellia sinensis): um potencial agente antitumoral em células de câncer de mama
title_full Extrato de chá verde (Camellia sinensis): um potencial agente antitumoral em células de câncer de mama
title_fullStr Extrato de chá verde (Camellia sinensis): um potencial agente antitumoral em células de câncer de mama
title_full_unstemmed Extrato de chá verde (Camellia sinensis): um potencial agente antitumoral em células de câncer de mama
title_sort Extrato de chá verde (Camellia sinensis): um potencial agente antitumoral em células de câncer de mama
author Santos, Ronimara Aparecida
author_facet Santos, Ronimara Aparecida
author_role author
dc.contributor.advisor1.fl_str_mv Costa, Danielly Cristiny Ferraz da
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/9617293991265023
dc.contributor.advisor-co1.fl_str_mv Daleprane, Júlio Beltrame
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/6027048461430935
dc.contributor.referee1.fl_str_mv Rangel, Luciana Pereira
dc.contributor.referee1Lattes.fl_str_mv http://lattes.cnpq.br/6987913099524130
dc.contributor.referee2.fl_str_mv Gimba, Etel Rodrigues Pereira
dc.contributor.referee2Lattes.fl_str_mv http://lattes.cnpq.br/8976892755529994
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/5223038433573741
dc.contributor.author.fl_str_mv Santos, Ronimara Aparecida
contributor_str_mv Costa, Danielly Cristiny Ferraz da
Daleprane, Júlio Beltrame
Rangel, Luciana Pereira
Gimba, Etel Rodrigues Pereira
dc.subject.eng.fl_str_mv Green tea
Poliphenols
Câncer
topic Green tea
Poliphenols
Câncer
Polifenóis
p53
Nutrição
Chá verde
Câncer
CNPQ::CIENCIAS DA SAUDE::NUTRICAO::DESNUTRICAO E DESENVOLVIMENTO FISIOLOGICO
dc.subject.por.fl_str_mv Polifenóis
p53
Nutrição
Chá verde
Câncer
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS DA SAUDE::NUTRICAO::DESNUTRICAO E DESENVOLVIMENTO FISIOLOGICO
description Green tea (GT), a beverage derived from Camellia sinensis, has been extensively studied for its antioxidant property, which is attributed to catechins. These phenolic compounds have been shown to inhibit carcinogenesis, but its mechanisms are still poorly understood. Additionally, most studies perform assays using isolated polyphenols from GT. We aimed to obtain a catechin-rich green tea extract (GTE) and to evaluate its antitumor potential on human breast cancer. Tea infusion was prepared on a ratio of 1 g: 40 mL of boiling distilled water using a combination of temperature and time (70, 75, 80, 85, 90, 95 and 100 ºC/5, 10, 15 min). Infusions were evaluated by polyphenols content (Folin Ciocalteu s reagent) and antioxidant potential (FRAP). Breast cancer cells (MDA-MB-231 and MCF-7) and non-tumoral cells (MCF-10A), were exposed to different concentrations of GTE (31.5 μg/mL to 1.0 mg/mL) during 24 h-48 h, and cell viability was assessed by Alamar Blue® and Trypan Blue assays. Viability was also acessed in the presence of p53 inhibitor, pifithrin-α. Immunocytochemistry and Western blotting analysis were performed to access the expression of p53. Our results demonstrated no influence of extraction condition in the total content of polyphenols and antioxidant potential of GT, and a new extract was obtained at 80 ºC/5 min, then freeze dried to be used in cell culture. After exposure for 24h, GTE already promoted a cytotoxic effect, reducing viability of both breast cancer cell lines (IC50 MDA-MB-231 = 138,8 μg/mL; IC50 MCF-7 = 324,5 μg/mL) without cytotoxic effect on normal cells (IC50 MCF-10A = 10097 μg/mL). In addition, GTE promotes an increase of p53 levels on treated MCF-7 cells, which express the wild-type form of the protein. Interestingly, an opposite phenomenon was evidenced in MDA-MB-231 cells, that express the mutated form of p53, in which protein levels seem to be lower in the presence of the extract. Our data suggests that GTE has anticarcinogenic potential on breast cancer and it is possible that this capacity might be related with the modulation of p53
publishDate 2018
dc.date.available.fl_str_mv 2018-12-10
dc.date.issued.fl_str_mv 2018-07-19
dc.date.accessioned.fl_str_mv 2021-01-05T16:42:19Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
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dc.identifier.citation.fl_str_mv SANTOS, Ronimara Aparecida. Extrato de chá verde (Camellia sinensis): um potencial agente antitumoral em células de câncer de mama. 2018. 22 f. Dissertação (Mestrado em Alimentação, Nutrição e Saúde) - Universidade do Estado do Rio de Janeiro, Rio de Janeiro, 2018.
dc.identifier.uri.fl_str_mv http://www.bdtd.uerj.br/handle/1/7244
identifier_str_mv SANTOS, Ronimara Aparecida. Extrato de chá verde (Camellia sinensis): um potencial agente antitumoral em células de câncer de mama. 2018. 22 f. Dissertação (Mestrado em Alimentação, Nutrição e Saúde) - Universidade do Estado do Rio de Janeiro, Rio de Janeiro, 2018.
url http://www.bdtd.uerj.br/handle/1/7244
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dc.publisher.country.fl_str_mv BR
dc.publisher.department.fl_str_mv Centro Biomédico::Instituto de Nutrição
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