O efeito da FNDC5 e Irisina em linhagens de células de câncer de mama MCF-7 e MDA-MB-231

Detalhes bibliográficos
Autor(a) principal: Fortes Junior, Pedro Florencio da Cunha
Data de Publicação: 2023
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da Universidade Federal do Espírito Santo (riUfes)
Texto Completo: http://repositorio.ufes.br/handle/10/17286
Resumo: Cancer is the second most prevalent disease and causes of deaths worldwide. Among the most prevalent cancers among women is breast cancer; It is estimated that 2.3 million new cases appear annually around the world. In this context, there is growing concern in the scientific field about the advancement of this disease. A myokine was discovered in 2012 that appears to reduce the proliferation of cancer cells, increasing the expression of pro-apoptotic proteins, decreasing the action of cell synthesis and proliferation mechanisms and other pro-inflammatory factors, in addition to regulating energy homeostasis. The objective of this work was to evaluate the effect of FNDC5 and Irisin molecules on human breast cancer lines. The recombinant molecules Irisin (glycosylated) and FNDC5 were used. The molecules were evaluated at concentrations of 0,625 nmol/L, 1,25 nmol/L, 2.5 nmol/L, 5 nmol/L, 10 nmol/L, 20 nmol/L and 40 nmol/L and the cells were treated48 hours. After this period, the MTT reagent was added at a concentration of 5 mg/mL, and incubation was carried out for 3 hours. After this period, the supernatant was discarded, 100 μl of DMSO reagent was added to all wells and, after 15 minutes, the absorbances were read at 540 nm. The estimated values of the 50% inhibitory concentration of cell proliferation (IC50) were calculated. The calculations were carried out using the GraphPad Prism software in version 8.0. Regarding the data from the MTT experiments, for normal data, one-way ANOVA with Dunnett's post hoc was used, and non-normal data were analyzed by Kruskall-Walli with Dunn's post hoc. The results showed that Irisin exerted cytotoxicity on the MDA-MB-231 strain at concentrations of 1.25 nmol/L (p<0.01), 5 nmol/L (p<0.01), 10 nmol/L (p< 0.0001), 20 nmol/L (p<0.05) and 40 nmol/L (p<0.01) with MCV values reaching 77.66%, 78.75%, 67.25%, 77, 83% and 75.5% respectively. In the MCF-7 line treated with Irisin, a reduction in viability was observed at a concentration of 2.5 nmol/L (p<0.05), a concentration at which the observed MCV was 83.58%. In turn, the FNDC5 molecule did not cause a statistically significant reduction in metabolic cell viability in the tested lines. Our data show that irisin can play an important role in treatment, with deleterious effects on the cell viability of cancer cells.
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spelling O efeito da FNDC5 e Irisina em linhagens de células de câncer de mama MCF-7 e MDA-MB-231title.alternativeCâncer de mamaMCF-7MDA-MB-231MiocionaFNDC5Irisinasubject.br-rjbnFarmacologia Bioquímica e MolecularCancer is the second most prevalent disease and causes of deaths worldwide. Among the most prevalent cancers among women is breast cancer; It is estimated that 2.3 million new cases appear annually around the world. In this context, there is growing concern in the scientific field about the advancement of this disease. A myokine was discovered in 2012 that appears to reduce the proliferation of cancer cells, increasing the expression of pro-apoptotic proteins, decreasing the action of cell synthesis and proliferation mechanisms and other pro-inflammatory factors, in addition to regulating energy homeostasis. The objective of this work was to evaluate the effect of FNDC5 and Irisin molecules on human breast cancer lines. The recombinant molecules Irisin (glycosylated) and FNDC5 were used. The molecules were evaluated at concentrations of 0,625 nmol/L, 1,25 nmol/L, 2.5 nmol/L, 5 nmol/L, 10 nmol/L, 20 nmol/L and 40 nmol/L and the cells were treated48 hours. After this period, the MTT reagent was added at a concentration of 5 mg/mL, and incubation was carried out for 3 hours. After this period, the supernatant was discarded, 100 μl of DMSO reagent was added to all wells and, after 15 minutes, the absorbances were read at 540 nm. The estimated values of the 50% inhibitory concentration of cell proliferation (IC50) were calculated. The calculations were carried out using the GraphPad Prism software in version 8.0. Regarding the data from the MTT experiments, for normal data, one-way ANOVA with Dunnett's post hoc was used, and non-normal data were analyzed by Kruskall-Walli with Dunn's post hoc. The results showed that Irisin exerted cytotoxicity on the MDA-MB-231 strain at concentrations of 1.25 nmol/L (p<0.01), 5 nmol/L (p<0.01), 10 nmol/L (p< 0.0001), 20 nmol/L (p<0.05) and 40 nmol/L (p<0.01) with MCV values reaching 77.66%, 78.75%, 67.25%, 77, 83% and 75.5% respectively. In the MCF-7 line treated with Irisin, a reduction in viability was observed at a concentration of 2.5 nmol/L (p<0.05), a concentration at which the observed MCV was 83.58%. In turn, the FNDC5 molecule did not cause a statistically significant reduction in metabolic cell viability in the tested lines. Our data show that irisin can play an important role in treatment, with deleterious effects on the cell viability of cancer cells.O câncer é a segunda doença mais prevalente e causadora de óbitos em todo o mundo. Entre os cânceres com maior prevalência entre as mulheres está o câncer de mama; estima-se que 2,3 milhões de novos casos surjam anualmente no mundo. Neste contexto há uma preocupação crescente no campo científico com avanço desta doença. Foi descoberta em 2012 uma miocina que parece diminuir a proliferação de células cancerosas, aumentando a expressão de proteínas pró-apoptóticas, diminuindo a ação de mecanismos de síntese e proliferação celular e outros fatores pró-inflamatórios, além de regular a homeostase energética. O objetivo deste trabalho foi avaliar o efeito das moléculas FNDC5 e Irisina em linhagens de câncer de mama humano. Foram utilizadas as moléculas recombinantes Irisina (glicosilada) e FNDC5. As moléculas foram avaliadas nas concentrações de 0,625 nmol/L, 1,25 nmol/L, 2,5 nmol/L, 5 nmol/L, 10 nmol/L, 20 nmol/L e 40 nmol/L e as células foram tratadas 48 horas. Após este período foi adicionado o reagente MTT na concentração de 5mg/mL, sendo feita a incubação por 3 horas. Após este período foi descartado o sobrenadante, sendo adicionados 100 μl do reagente DMSO em todos os poços e, após 15 minutos, foi feita a leitura das absorbâncias a 540 nm. Foi efetuado o cálculo estimado dos valores da concentração inibitória de 50% da proliferação celular (IC50). Os cálculos foram efetuados com a utilização do software GraphPad Prism na versão 8.0. Em relação aos dados dos experimentos de MTT, para dados normais foi utilizada ANOVA de uma via com post hoc de Dunnett, e os dados não normais foram analisados por Kruskall-Walli com post hoc de Dunn. Os resultados mostraram que a Irisina exerceu citotoxicidade na linhagem MDA-MB-231 nas concentrações 1,25 nmol/L (p<0,01), 5 nmol/L (p<0,01), 10 nmol/L (p<0,0001), 20 nmol/L (p<0,05) e 40 nmol/L (p<0,01) com valores de VCM atingindo 77,66%, 78,75%, 67,25%, 77,83% e 75,5% respectivamente. Na linhagem MCF-7 tratada com Irisina, foi observada redução na viabilidade com a concentração de 2,5 nmol/L (p<0,05), concentração na qual a VCM observada foi de 83,58%. Por sua vez, a molécula FNDC5 não provocou redução estatisticamente significante na viabilidade celular metabólica nas linhagens testadas. Nossos dados mostram que a irisina pode ter um papel importante no tratamento, com efeitos deletérios sobre a viabilidade celular das células cancerosas.Fundação de Amparo à Pesquisa do Espírito Santo (FAPES)Universidade Federal do Espírito SantoBRMestrado em BioquímicaCentro de Ciências da SaúdeUFESPrograma de Pós-Graduação em BioquímicaDaltoe, Renata Dalmaschiohttps://orcid.org/0000000285710428http://lattes.cnpq.br/0931389996547944https://orcid.org/0009-0007-4315-0315http://lattes.cnpq.br/5958617857750748Sousa, Nuno Manuel Frade dehttp://lattes.cnpq.br/0299657415152651Madeira, Klesia Pirolahttps://orcid.org/0000-0002-9442-9961http://lattes.cnpq.br/2179692838971480Gonçalves, Juliana Barbosa Coitinhohttps://orcid.org/000000025892050Xhttp://lattes.cnpq.br/3448669742301744Fortes Junior, Pedro Florencio da Cunha2024-05-30T01:43:03Z2024-05-30T01:43:03Z2023-09-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisTextapplication/pdfhttp://repositorio.ufes.br/handle/10/17286porinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da Universidade Federal do Espírito Santo (riUfes)instname:Universidade Federal do Espírito Santo (UFES)instacron:UFES2024-10-14T08:15:31Zoai:repositorio.ufes.br:10/17286Repositório InstitucionalPUBhttp://repositorio.ufes.br/oai/requestopendoar:21082024-10-14T08:15:31Repositório Institucional da Universidade Federal do Espírito Santo (riUfes) - Universidade Federal do Espírito Santo (UFES)false
dc.title.none.fl_str_mv O efeito da FNDC5 e Irisina em linhagens de células de câncer de mama MCF-7 e MDA-MB-231
title.alternative
title O efeito da FNDC5 e Irisina em linhagens de células de câncer de mama MCF-7 e MDA-MB-231
spellingShingle O efeito da FNDC5 e Irisina em linhagens de células de câncer de mama MCF-7 e MDA-MB-231
Fortes Junior, Pedro Florencio da Cunha
Câncer de mama
MCF-7
MDA-MB-231
Miociona
FNDC5
Irisina
subject.br-rjbn
Farmacologia Bioquímica e Molecular
title_short O efeito da FNDC5 e Irisina em linhagens de células de câncer de mama MCF-7 e MDA-MB-231
title_full O efeito da FNDC5 e Irisina em linhagens de células de câncer de mama MCF-7 e MDA-MB-231
title_fullStr O efeito da FNDC5 e Irisina em linhagens de células de câncer de mama MCF-7 e MDA-MB-231
title_full_unstemmed O efeito da FNDC5 e Irisina em linhagens de células de câncer de mama MCF-7 e MDA-MB-231
title_sort O efeito da FNDC5 e Irisina em linhagens de células de câncer de mama MCF-7 e MDA-MB-231
author Fortes Junior, Pedro Florencio da Cunha
author_facet Fortes Junior, Pedro Florencio da Cunha
author_role author
dc.contributor.none.fl_str_mv Daltoe, Renata Dalmaschio
https://orcid.org/0000000285710428
http://lattes.cnpq.br/0931389996547944
https://orcid.org/0009-0007-4315-0315
http://lattes.cnpq.br/5958617857750748
Sousa, Nuno Manuel Frade de
http://lattes.cnpq.br/0299657415152651
Madeira, Klesia Pirola
https://orcid.org/0000-0002-9442-9961
http://lattes.cnpq.br/2179692838971480
Gonçalves, Juliana Barbosa Coitinho
https://orcid.org/000000025892050X
http://lattes.cnpq.br/3448669742301744
dc.contributor.author.fl_str_mv Fortes Junior, Pedro Florencio da Cunha
dc.subject.por.fl_str_mv Câncer de mama
MCF-7
MDA-MB-231
Miociona
FNDC5
Irisina
subject.br-rjbn
Farmacologia Bioquímica e Molecular
topic Câncer de mama
MCF-7
MDA-MB-231
Miociona
FNDC5
Irisina
subject.br-rjbn
Farmacologia Bioquímica e Molecular
description Cancer is the second most prevalent disease and causes of deaths worldwide. Among the most prevalent cancers among women is breast cancer; It is estimated that 2.3 million new cases appear annually around the world. In this context, there is growing concern in the scientific field about the advancement of this disease. A myokine was discovered in 2012 that appears to reduce the proliferation of cancer cells, increasing the expression of pro-apoptotic proteins, decreasing the action of cell synthesis and proliferation mechanisms and other pro-inflammatory factors, in addition to regulating energy homeostasis. The objective of this work was to evaluate the effect of FNDC5 and Irisin molecules on human breast cancer lines. The recombinant molecules Irisin (glycosylated) and FNDC5 were used. The molecules were evaluated at concentrations of 0,625 nmol/L, 1,25 nmol/L, 2.5 nmol/L, 5 nmol/L, 10 nmol/L, 20 nmol/L and 40 nmol/L and the cells were treated48 hours. After this period, the MTT reagent was added at a concentration of 5 mg/mL, and incubation was carried out for 3 hours. After this period, the supernatant was discarded, 100 μl of DMSO reagent was added to all wells and, after 15 minutes, the absorbances were read at 540 nm. The estimated values of the 50% inhibitory concentration of cell proliferation (IC50) were calculated. The calculations were carried out using the GraphPad Prism software in version 8.0. Regarding the data from the MTT experiments, for normal data, one-way ANOVA with Dunnett's post hoc was used, and non-normal data were analyzed by Kruskall-Walli with Dunn's post hoc. The results showed that Irisin exerted cytotoxicity on the MDA-MB-231 strain at concentrations of 1.25 nmol/L (p<0.01), 5 nmol/L (p<0.01), 10 nmol/L (p< 0.0001), 20 nmol/L (p<0.05) and 40 nmol/L (p<0.01) with MCV values reaching 77.66%, 78.75%, 67.25%, 77, 83% and 75.5% respectively. In the MCF-7 line treated with Irisin, a reduction in viability was observed at a concentration of 2.5 nmol/L (p<0.05), a concentration at which the observed MCV was 83.58%. In turn, the FNDC5 molecule did not cause a statistically significant reduction in metabolic cell viability in the tested lines. Our data show that irisin can play an important role in treatment, with deleterious effects on the cell viability of cancer cells.
publishDate 2023
dc.date.none.fl_str_mv 2023-09-01
2024-05-30T01:43:03Z
2024-05-30T01:43:03Z
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.publisher.none.fl_str_mv Universidade Federal do Espírito Santo
BR
Mestrado em Bioquímica
Centro de Ciências da Saúde
UFES
Programa de Pós-Graduação em Bioquímica
publisher.none.fl_str_mv Universidade Federal do Espírito Santo
BR
Mestrado em Bioquímica
Centro de Ciências da Saúde
UFES
Programa de Pós-Graduação em Bioquímica
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