Atividade antitumoral da berberina em células de neuroblastoma SH-SY5Y

Detalhes bibliográficos
Autor(a) principal: Lenz, Luana Suéling
Data de Publicação: 2017
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Manancial - Repositório Digital da UFSM
Texto Completo: http://repositorio.ufsm.br/handle/1/20950
Resumo: Neuroblastoma is the most common tumor found in the first year of life and represent 6% of the all childhood cancers. There is few options of treatment and the mortality is close to 21%, or exceeds 50% between the high risk cases the mortality. Berberine (BBR) is an alkaloid used in traditional Chinese medicine for a long time and several studies have proved a diversity of pharmacological activities from this compound, among them antitumoral. Therefore, this molecule may have potential to the treatment of neuroblastoma. The aim of this study was to evaluate whether BBR has antitumoral activity in SH-SY5Y neuroblastoma cell lineage, as well as, investigate if the mechanism trigger is through ERO induction, AMPK and p53 activation. First of all, it was performed a cell death assay with several different concentration of BBR. From this data, we identify the minor concentration able to lead cell to death, which was used in the next experiments. In cells treated or not with BBR and/or an AMPK inhibitor (Compound C), it was determined the apoptotic index and cell cycle measure, as well as labeling with specific antibody to quantify the phosphor-p53 and caspase 3. In the same way, the migration and forming colony ability, ERO levels, activity of antioxidant enzymes and oxidative damage on DNA was detected. Still, it was done q-PCR to quantify the expression of p53, bax, bcl-2, caspase-3 and antioxidant enzymes genes. Our results prove that BBR do not alter cell cycle in cell line studied, unlike the results from other researches, but increase the apoptotic cells and the levels of p-p53 independent of AMPK activity, besides causing an increase in ERO levels and catalase activity. Also, it was detected a decrease in migratory and forming colony ability in the cells treated. This study suggest that BBR have therapeutic potential as an adjuvant in neuroblastoma treatment.
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spelling Atividade antitumoral da berberina em células de neuroblastoma SH-SY5YAntimural activity of berberine on neuroblastoma SH-SY5Y cellNeuroblastomaBerberinaEstresse oxidativoBerberineNeuroblastomaOxidative stressCNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICANeuroblastoma is the most common tumor found in the first year of life and represent 6% of the all childhood cancers. There is few options of treatment and the mortality is close to 21%, or exceeds 50% between the high risk cases the mortality. Berberine (BBR) is an alkaloid used in traditional Chinese medicine for a long time and several studies have proved a diversity of pharmacological activities from this compound, among them antitumoral. Therefore, this molecule may have potential to the treatment of neuroblastoma. The aim of this study was to evaluate whether BBR has antitumoral activity in SH-SY5Y neuroblastoma cell lineage, as well as, investigate if the mechanism trigger is through ERO induction, AMPK and p53 activation. First of all, it was performed a cell death assay with several different concentration of BBR. From this data, we identify the minor concentration able to lead cell to death, which was used in the next experiments. In cells treated or not with BBR and/or an AMPK inhibitor (Compound C), it was determined the apoptotic index and cell cycle measure, as well as labeling with specific antibody to quantify the phosphor-p53 and caspase 3. In the same way, the migration and forming colony ability, ERO levels, activity of antioxidant enzymes and oxidative damage on DNA was detected. Still, it was done q-PCR to quantify the expression of p53, bax, bcl-2, caspase-3 and antioxidant enzymes genes. Our results prove that BBR do not alter cell cycle in cell line studied, unlike the results from other researches, but increase the apoptotic cells and the levels of p-p53 independent of AMPK activity, besides causing an increase in ERO levels and catalase activity. Also, it was detected a decrease in migratory and forming colony ability in the cells treated. This study suggest that BBR have therapeutic potential as an adjuvant in neuroblastoma treatment.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESNeuroblastoma é o tumor mais comum encontrado no primeiro ano de vida e representa 6% do total de cânceres infantis. Existem poucas opções de tratamento e a mortalidade chega a 21%, entre os casos de alto risco a mortalidade ultrapassa os 50%. A berberina é um alcaloide utilizado na medicina tradicional chinesa por muito tempo e diversos estudos têm comprovado atividades farmacológicas variadas desse composto, entre elas a antitumoral. Sendo assim, esta molécula pode ter potencial no tratamento do neuroblastoma. O objetivo desse trabalho foi avaliar se a BBR possui ação anti-tumoral na linhagem de neuroblastoma SH-SY5Y, bem como, investigar seu possível mecanismo através da via das espécies reativas de oxigênio (ERO), proteína quinase ativada por AMP (AMPK) e p53. Inicialmente, foi realizado um ensaio de morte celular com diferentes concentrações de BBR. A partir desses dados, identificamos a menor concentração capaz de levar as células a morte, a qual foi usada nos testes posteriores. Em células não tratadas ou tratadas com BBR e/ou um inibidor de AMPK (composto C), foi determinado os índices de apoptose e medidas do ciclo celular, bem como marcação com anticorpos específicos para quantificar a fosfo-p53 (Ser-15) e caspase 3. Paralelamente, a capacidade de migração e de formação de colônias, os níveis de ERO, atividade das enzimas antioxidantes e danos oxidativos ao DNA foram avaliados. Ainda, foi realizado q-PCR para quantificar a expressão dos genes p53, bax, bcl-2, caspase-3 e enzimas antioxidantes. Nossos resultados demonstram que a BBR não altera o ciclo celular nas células estudadas, diferente do que é encontrado descrito em outros estudos, mas provoca um aumento na quantidade de células apoptóticas e dos níveis de p-p53 independente da atividade da AMPK, além de provocar aumento das ERO e da atividade da enzima catalase. Também, houve uma redução na capacidade migratória e de formação de colônias nas células tratadas. Sugere-se com este estudo que a BBR tem potencial terapêutico como adjuvante no tratamento do neuroblastoma.Universidade Federal de Santa MariaBrasilBioquímicaUFSMPrograma de Pós-Graduação em Ciências Biológicas: Bioquímica ToxicológicaCentro de Ciências Naturais e ExatasAndrade, Cinthia Melazzo dehttp://lattes.cnpq.br/2886709251370905Pillat, Micheli MainardiSagrillo, Michele RoratoBauermann, LilianeLenz, Luana Suéling2021-05-21T11:17:38Z2021-05-21T11:17:38Z2017-02-23info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://repositorio.ufsm.br/handle/1/20950porAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2021-09-29T19:40:35Zoai:repositorio.ufsm.br:1/20950Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2021-09-29T19:40:35Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false
dc.title.none.fl_str_mv Atividade antitumoral da berberina em células de neuroblastoma SH-SY5Y
Antimural activity of berberine on neuroblastoma SH-SY5Y cell
title Atividade antitumoral da berberina em células de neuroblastoma SH-SY5Y
spellingShingle Atividade antitumoral da berberina em células de neuroblastoma SH-SY5Y
Lenz, Luana Suéling
Neuroblastoma
Berberina
Estresse oxidativo
Berberine
Neuroblastoma
Oxidative stress
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA
title_short Atividade antitumoral da berberina em células de neuroblastoma SH-SY5Y
title_full Atividade antitumoral da berberina em células de neuroblastoma SH-SY5Y
title_fullStr Atividade antitumoral da berberina em células de neuroblastoma SH-SY5Y
title_full_unstemmed Atividade antitumoral da berberina em células de neuroblastoma SH-SY5Y
title_sort Atividade antitumoral da berberina em células de neuroblastoma SH-SY5Y
author Lenz, Luana Suéling
author_facet Lenz, Luana Suéling
author_role author
dc.contributor.none.fl_str_mv Andrade, Cinthia Melazzo de
http://lattes.cnpq.br/2886709251370905
Pillat, Micheli Mainardi
Sagrillo, Michele Rorato
Bauermann, Liliane
dc.contributor.author.fl_str_mv Lenz, Luana Suéling
dc.subject.por.fl_str_mv Neuroblastoma
Berberina
Estresse oxidativo
Berberine
Neuroblastoma
Oxidative stress
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA
topic Neuroblastoma
Berberina
Estresse oxidativo
Berberine
Neuroblastoma
Oxidative stress
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA
description Neuroblastoma is the most common tumor found in the first year of life and represent 6% of the all childhood cancers. There is few options of treatment and the mortality is close to 21%, or exceeds 50% between the high risk cases the mortality. Berberine (BBR) is an alkaloid used in traditional Chinese medicine for a long time and several studies have proved a diversity of pharmacological activities from this compound, among them antitumoral. Therefore, this molecule may have potential to the treatment of neuroblastoma. The aim of this study was to evaluate whether BBR has antitumoral activity in SH-SY5Y neuroblastoma cell lineage, as well as, investigate if the mechanism trigger is through ERO induction, AMPK and p53 activation. First of all, it was performed a cell death assay with several different concentration of BBR. From this data, we identify the minor concentration able to lead cell to death, which was used in the next experiments. In cells treated or not with BBR and/or an AMPK inhibitor (Compound C), it was determined the apoptotic index and cell cycle measure, as well as labeling with specific antibody to quantify the phosphor-p53 and caspase 3. In the same way, the migration and forming colony ability, ERO levels, activity of antioxidant enzymes and oxidative damage on DNA was detected. Still, it was done q-PCR to quantify the expression of p53, bax, bcl-2, caspase-3 and antioxidant enzymes genes. Our results prove that BBR do not alter cell cycle in cell line studied, unlike the results from other researches, but increase the apoptotic cells and the levels of p-p53 independent of AMPK activity, besides causing an increase in ERO levels and catalase activity. Also, it was detected a decrease in migratory and forming colony ability in the cells treated. This study suggest that BBR have therapeutic potential as an adjuvant in neuroblastoma treatment.
publishDate 2017
dc.date.none.fl_str_mv 2017-02-23
2021-05-21T11:17:38Z
2021-05-21T11:17:38Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://repositorio.ufsm.br/handle/1/20950
url http://repositorio.ufsm.br/handle/1/20950
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Santa Maria
Brasil
Bioquímica
UFSM
Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica
Centro de Ciências Naturais e Exatas
publisher.none.fl_str_mv Universidade Federal de Santa Maria
Brasil
Bioquímica
UFSM
Programa de Pós-Graduação em Ciências Biológicas: Bioquímica Toxicológica
Centro de Ciências Naturais e Exatas
dc.source.none.fl_str_mv reponame:Manancial - Repositório Digital da UFSM
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instacron:UFSM
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institution UFSM
reponame_str Manancial - Repositório Digital da UFSM
collection Manancial - Repositório Digital da UFSM
repository.name.fl_str_mv Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)
repository.mail.fl_str_mv atendimento.sib@ufsm.br||tedebc@gmail.com
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