Efeitos de uma desintegrina RGD recombinante de Rhinocerophis alternatus (Bothrops alternatus) na migração de células normais e tumorais

Detalhes bibliográficos
Autor(a) principal: Montenegro, Cyntia de Freitas
Data de Publicação: 2011
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da UFSCAR
Texto Completo: https://repositorio.ufscar.br/handle/ufscar/1342
Resumo: Cell migration, invasion of the surrounding tissues, blood vessels and proliferation are cellular processes involved in metastases. These events are mediated by integrins, adhesion receptors present on cell surfaces and responsible for the extracellular matrix and cells binding. Disintegrins, snake venoms polypeptides, are known to inhibit integrins action, and thus metastatic events such as cell migration. They also have influence on metallopeptidases activity, enzymes that cleave ECM components facilitating tumor cells invasion. DisBa-01 is a recombinant RGD disintegrin from Rhinocerophis alternatus (Bothrops alternatus) venom gland. In previous studies we have demonstrated that DisBa-01 binds to _v_3 integrin with high affinity and has an anti-metastatic activity. Therefore the aim of this study was to investigate the effect of DisBa-01 on migration and MMPs activities in fibroblasts and cell lines of breast cancer, MDA-MB-231, prostate, DU-145 and murine melanoma, B16F10. Transwell migration tests, wound healing and gelatin gel zymography were performed using DisBa-01 at concentrations of 10, 100, 500 and 1000 nM. Transwell migration was significantly inhibited in fibroblasts, MDA-MB-231 and DU-145 but had no effect on B16F10 cells, which had fibroblast conditioned medium as chemoattractant. In wound healing assays only DU-145 migration in a 1000nM concentration was significantly inhibited. Finally, zymography was performed using wound healing assay conditioned medium to analyze the presence of MMPs 2 and 9. In fibroblasts and B16F10 cells MMP-2 and MMPs 2 and 9 were decreased respectively. On the other hand, MDA-MB-231 cells showed increased MMP-9 activity, in all concentrations tested. Given the results, DisBa-01 can be used to build models for possible drugs rational design that may act in anti-metastatic therapy, in addition to its use as a research tool to elucidate integrins related mechanisms.
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spelling Montenegro, Cyntia de FreitasAraújo, Heloísa Sobreiro Selistre dehttp://genos.cnpq.br:12010/dwlattes/owa/prc_imp_cv_int?f_cod=K4787059Y2http://lattes.cnpq.br/1510751976021672e39baeab-9ced-4188-ae4a-a5fea86125f82016-06-02T19:22:56Z2012-09-052016-06-02T19:22:56Z2011-08-03MONTENEGRO, Cyntia de Freitas. Efeitos de uma desintegrina RGD recombinante de Rhinocerophis alternatus (Bothrops alternatus) na migração de células normais e tumorais. 2011. 84 f. Dissertação (Mestrado em Ciências Biológicas) - Universidade Federal de São Carlos, São Carlos, 2011.https://repositorio.ufscar.br/handle/ufscar/1342Cell migration, invasion of the surrounding tissues, blood vessels and proliferation are cellular processes involved in metastases. These events are mediated by integrins, adhesion receptors present on cell surfaces and responsible for the extracellular matrix and cells binding. Disintegrins, snake venoms polypeptides, are known to inhibit integrins action, and thus metastatic events such as cell migration. They also have influence on metallopeptidases activity, enzymes that cleave ECM components facilitating tumor cells invasion. DisBa-01 is a recombinant RGD disintegrin from Rhinocerophis alternatus (Bothrops alternatus) venom gland. In previous studies we have demonstrated that DisBa-01 binds to _v_3 integrin with high affinity and has an anti-metastatic activity. Therefore the aim of this study was to investigate the effect of DisBa-01 on migration and MMPs activities in fibroblasts and cell lines of breast cancer, MDA-MB-231, prostate, DU-145 and murine melanoma, B16F10. Transwell migration tests, wound healing and gelatin gel zymography were performed using DisBa-01 at concentrations of 10, 100, 500 and 1000 nM. Transwell migration was significantly inhibited in fibroblasts, MDA-MB-231 and DU-145 but had no effect on B16F10 cells, which had fibroblast conditioned medium as chemoattractant. In wound healing assays only DU-145 migration in a 1000nM concentration was significantly inhibited. Finally, zymography was performed using wound healing assay conditioned medium to analyze the presence of MMPs 2 and 9. In fibroblasts and B16F10 cells MMP-2 and MMPs 2 and 9 were decreased respectively. On the other hand, MDA-MB-231 cells showed increased MMP-9 activity, in all concentrations tested. Given the results, DisBa-01 can be used to build models for possible drugs rational design that may act in anti-metastatic therapy, in addition to its use as a research tool to elucidate integrins related mechanisms.Uma série de processos celulares está envolvida na metástase, entre eles a migração e invasão celular dos tecidos vizinhos, vasos sanguíneos e proliferação. Esses eventos são mediados por integrinas, receptores de adesão, presentes nas superfícies celulares, responsáveis pela ligação das células com a matriz extracelular. As desintegrinas, polipeptídeos provenientes de venenos de serpentes, são conhecidos por inibir a ação das integrinas e, consequentemente, eventos como a migração, que ocorrem durante a metástase. Possuem ainda, influência sobre a atividade das metalopeptidases (MMPs), enzimas que clivam a MEC facilitando a invasão das células tumorais. A DisBa-01 é uma desintegrina recombinante que possui motivo RGD e é proveniente do veneno da Rhinocerophis alternatus (Bothrops alternatus). Estudos anteriores comprovaram sua alta afinidade pela integrina _v_3 e atividade anti-metastática in vivo. Portanto, o objetivo desse estudo foi investigar a ação da DisBa-01 sobre a migração e atividade das MMPs de fibroblastos e das linhagens de câncer de mama, MDA-MB-231, próstata, DU-145 e melanoma murino, B16F10. Foram realizados ensaios de migração em transwell, wound healing, e zimografia em gel de poliacrilamida acrescido de gelatina utilizando a DisBa-01 nas concentrações de 10, 100, 500 e 1000nM. A inibição da migração em transwell foi significativa nos fibroblastos, MDA-MB-231 e DU-145, porém não houve efeito sobre a linhagem B16F10, que teve como quimioatraente o meio condicionado de fibroblastos. Nos ensaios de wound healing apenas a concentração de 1000 nM da DU-145 foi inibida de forma significativa. Finalmente, a zimografia foi realizada utilizando meio condicionado de ensaios wound healing para analisar a presença das MMPs 2 e 9. Nos fibroblastos e B16F10 a atividade da MMP-2 e as MMPs 2 e 9 foram inibidas respectivamente. Enquanto a linhagem MDA-MB-231 teve a secreção da MMP-9 estimulada, em todas as concentrações, de forma significativa. Tendo em vista os resultados, a DisBa-01 pode ser utilizada na construção de modelos para o desenho racional de possíveis fármacos que atuem na terapia anti-metastática, além da sua utilização como instrumento de pesquisa para elucidação de mecanismos relacionados as integrinas.Universidade Federal de Sao Carlosapplication/pdfporUniversidade Federal de São CarlosPrograma Interinstitucional de Pós-Graduação em Ciências Fisiológicas - PIPGCFUFSCarBRCâncerCélulas migraçãoDisBa-01DesintegrinaMetástaseRhinocerophis alternatusCIENCIAS BIOLOGICAS::FISIOLOGIAEfeitos de uma desintegrina RGD recombinante de Rhinocerophis alternatus (Bothrops alternatus) na migração de células normais e tumoraisinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesis-1-1b61211db-d955-45b7-886e-a0cefb89980finfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINAL4455.pdfapplication/pdf6484329https://repositorio.ufscar.br/bitstream/ufscar/1342/1/4455.pdf0fee43406bc059da2f0e202d0d04a19eMD51TEXT4455.pdf.txt4455.pdf.txtExtracted texttext/plain0https://repositorio.ufscar.br/bitstream/ufscar/1342/2/4455.pdf.txtd41d8cd98f00b204e9800998ecf8427eMD52THUMBNAIL4455.pdf.jpg4455.pdf.jpgIM Thumbnailimage/jpeg7464https://repositorio.ufscar.br/bitstream/ufscar/1342/3/4455.pdf.jpg3458416c8dd435dcca2766a506a0ce8cMD53ufscar/13422023-09-18 18:31:48.665oai:repositorio.ufscar.br:ufscar/1342Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222023-09-18T18:31:48Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false
dc.title.por.fl_str_mv Efeitos de uma desintegrina RGD recombinante de Rhinocerophis alternatus (Bothrops alternatus) na migração de células normais e tumorais
title Efeitos de uma desintegrina RGD recombinante de Rhinocerophis alternatus (Bothrops alternatus) na migração de células normais e tumorais
spellingShingle Efeitos de uma desintegrina RGD recombinante de Rhinocerophis alternatus (Bothrops alternatus) na migração de células normais e tumorais
Montenegro, Cyntia de Freitas
Câncer
Células migração
DisBa-01
Desintegrina
Metástase
Rhinocerophis alternatus
CIENCIAS BIOLOGICAS::FISIOLOGIA
title_short Efeitos de uma desintegrina RGD recombinante de Rhinocerophis alternatus (Bothrops alternatus) na migração de células normais e tumorais
title_full Efeitos de uma desintegrina RGD recombinante de Rhinocerophis alternatus (Bothrops alternatus) na migração de células normais e tumorais
title_fullStr Efeitos de uma desintegrina RGD recombinante de Rhinocerophis alternatus (Bothrops alternatus) na migração de células normais e tumorais
title_full_unstemmed Efeitos de uma desintegrina RGD recombinante de Rhinocerophis alternatus (Bothrops alternatus) na migração de células normais e tumorais
title_sort Efeitos de uma desintegrina RGD recombinante de Rhinocerophis alternatus (Bothrops alternatus) na migração de células normais e tumorais
author Montenegro, Cyntia de Freitas
author_facet Montenegro, Cyntia de Freitas
author_role author
dc.contributor.authorlattes.por.fl_str_mv http://lattes.cnpq.br/1510751976021672
dc.contributor.author.fl_str_mv Montenegro, Cyntia de Freitas
dc.contributor.advisor1.fl_str_mv Araújo, Heloísa Sobreiro Selistre de
dc.contributor.advisor1Lattes.fl_str_mv http://genos.cnpq.br:12010/dwlattes/owa/prc_imp_cv_int?f_cod=K4787059Y2
dc.contributor.authorID.fl_str_mv e39baeab-9ced-4188-ae4a-a5fea86125f8
contributor_str_mv Araújo, Heloísa Sobreiro Selistre de
dc.subject.por.fl_str_mv Câncer
Células migração
DisBa-01
Desintegrina
Metástase
topic Câncer
Células migração
DisBa-01
Desintegrina
Metástase
Rhinocerophis alternatus
CIENCIAS BIOLOGICAS::FISIOLOGIA
dc.subject.lat.fl_str_mv Rhinocerophis alternatus
dc.subject.cnpq.fl_str_mv CIENCIAS BIOLOGICAS::FISIOLOGIA
description Cell migration, invasion of the surrounding tissues, blood vessels and proliferation are cellular processes involved in metastases. These events are mediated by integrins, adhesion receptors present on cell surfaces and responsible for the extracellular matrix and cells binding. Disintegrins, snake venoms polypeptides, are known to inhibit integrins action, and thus metastatic events such as cell migration. They also have influence on metallopeptidases activity, enzymes that cleave ECM components facilitating tumor cells invasion. DisBa-01 is a recombinant RGD disintegrin from Rhinocerophis alternatus (Bothrops alternatus) venom gland. In previous studies we have demonstrated that DisBa-01 binds to _v_3 integrin with high affinity and has an anti-metastatic activity. Therefore the aim of this study was to investigate the effect of DisBa-01 on migration and MMPs activities in fibroblasts and cell lines of breast cancer, MDA-MB-231, prostate, DU-145 and murine melanoma, B16F10. Transwell migration tests, wound healing and gelatin gel zymography were performed using DisBa-01 at concentrations of 10, 100, 500 and 1000 nM. Transwell migration was significantly inhibited in fibroblasts, MDA-MB-231 and DU-145 but had no effect on B16F10 cells, which had fibroblast conditioned medium as chemoattractant. In wound healing assays only DU-145 migration in a 1000nM concentration was significantly inhibited. Finally, zymography was performed using wound healing assay conditioned medium to analyze the presence of MMPs 2 and 9. In fibroblasts and B16F10 cells MMP-2 and MMPs 2 and 9 were decreased respectively. On the other hand, MDA-MB-231 cells showed increased MMP-9 activity, in all concentrations tested. Given the results, DisBa-01 can be used to build models for possible drugs rational design that may act in anti-metastatic therapy, in addition to its use as a research tool to elucidate integrins related mechanisms.
publishDate 2011
dc.date.issued.fl_str_mv 2011-08-03
dc.date.available.fl_str_mv 2012-09-05
2016-06-02T19:22:56Z
dc.date.accessioned.fl_str_mv 2016-06-02T19:22:56Z
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dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
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dc.identifier.citation.fl_str_mv MONTENEGRO, Cyntia de Freitas. Efeitos de uma desintegrina RGD recombinante de Rhinocerophis alternatus (Bothrops alternatus) na migração de células normais e tumorais. 2011. 84 f. Dissertação (Mestrado em Ciências Biológicas) - Universidade Federal de São Carlos, São Carlos, 2011.
dc.identifier.uri.fl_str_mv https://repositorio.ufscar.br/handle/ufscar/1342
identifier_str_mv MONTENEGRO, Cyntia de Freitas. Efeitos de uma desintegrina RGD recombinante de Rhinocerophis alternatus (Bothrops alternatus) na migração de células normais e tumorais. 2011. 84 f. Dissertação (Mestrado em Ciências Biológicas) - Universidade Federal de São Carlos, São Carlos, 2011.
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