Atividade antitumoral da berberina em células de neuroblastoma SH-SY5Y
Autor(a) principal: | |
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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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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 instname:Universidade Federal de Santa Maria (UFSM) instacron:UFSM |
instname_str |
Universidade Federal de Santa Maria (UFSM) |
instacron_str |
UFSM |
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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1805922085570084864 |