Tumor mamário canino: estudo in vitro, imunomarcação e ação da doxorrubicina

Detalhes bibliográficos
Autor(a) principal: Lima, Silmara Sanae Sakamoto de [UNESP]
Data de Publicação: 2014
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://hdl.handle.net/11449/128081
http://www.athena.biblioteca.unesp.br/exlibris/bd/cathedra/16-09-2015/000850064.pdf
Resumo: Canine mammary tumors represent a high casuistry in veterinary oncology and few studies with cell culture are being employed, mainly with drug application. The purpose was to establish and standardize the cell culture of canine mammary tumors for in vitro efficacy of doxorubicin and to correlate the immunostaining of vimentin, cytokeratin, p53 and HER-2, in paraffin sections of original tumor and the cell cultures derivated. Benign mixed tumor, complex carcinoma, simple carcinoma and espinocelular carcinoma were cultivated from explants and the cells originated were incubated with doxorubicin in different concentrations (control-0.25-0.50-0.75-1.0-2.0μM). The percentual of viable cells were determined by the Trypan blue dye exclusion test and for immunoexpression, the immunohistochemistry was performed for the primary tumor and immunofluorescence, for the cells. A total of 39 tumor samples were collected and, 40% were cultivated up to fifth passages. A different imunnoexpression was observed between methodologies with higher expression at vimentin, p53 and HER-2 groups. It was observed that doxorubicin presented an elevated efficacy in canine mammary cell culture but without difference among the histological groups and each sample presented an individual responsiveness for the same treatment. So, it can be concluded that it was possible to realize a primary cell culture with a simple culture medium formulation; gene expression profiles in tumor tissue and cell culture derived from those tumors are better designated than immunoexpression tests and higher the concentration of doxorubicin, lower the cell viability
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spelling Tumor mamário canino: estudo in vitro, imunomarcação e ação da doxorrubicinaCãoMamas - CancerQuimioterapia veterinariaDoxorrubicinaCelulas - Cultura e meios de culturaDog diseasesCanine mammary tumors represent a high casuistry in veterinary oncology and few studies with cell culture are being employed, mainly with drug application. The purpose was to establish and standardize the cell culture of canine mammary tumors for in vitro efficacy of doxorubicin and to correlate the immunostaining of vimentin, cytokeratin, p53 and HER-2, in paraffin sections of original tumor and the cell cultures derivated. Benign mixed tumor, complex carcinoma, simple carcinoma and espinocelular carcinoma were cultivated from explants and the cells originated were incubated with doxorubicin in different concentrations (control-0.25-0.50-0.75-1.0-2.0μM). The percentual of viable cells were determined by the Trypan blue dye exclusion test and for immunoexpression, the immunohistochemistry was performed for the primary tumor and immunofluorescence, for the cells. A total of 39 tumor samples were collected and, 40% were cultivated up to fifth passages. A different imunnoexpression was observed between methodologies with higher expression at vimentin, p53 and HER-2 groups. It was observed that doxorubicin presented an elevated efficacy in canine mammary cell culture but without difference among the histological groups and each sample presented an individual responsiveness for the same treatment. So, it can be concluded that it was possible to realize a primary cell culture with a simple culture medium formulation; gene expression profiles in tumor tissue and cell culture derived from those tumors are better designated than immunoexpression tests and higher the concentration of doxorubicin, lower the cell viabilityA neoplasia mamária canina representa uma das maiores casuísticas na oncologia veterinária e o emprego de seus respectivos cultivos celulares são importantes, principalmente, na área terapêutica. Assim, o objetivo do presente estudo foi padronizar e estabelecer cultivos celulares de tumores mamários caninos para testes in vitro com doxorrubicina e, correlacionar imunomarcações com vimentina, citoqueratina, p53 e HER-2 no tumor original e nas células do seu cultivo. Tumor misto benigno e os carcinomas complexo, simples e espinocelular foram cultivados a partir de explantes com posterior exposição das células à doxorrubicina (controle-0,25-0,50-0,75-1,0-2,0μM). A viabilidade celular foi mensurada pelo azul de Tripan e os testes com imunomarcadores para cada espécie tumoral foram por imuno-histoquímica e, no cultivo, pela imunofluorescência. Do total de 39 amostras, 40% foram cultivadas até a quinta ou mais passagens. As imunomarcações não foram expressas de forma similar entre as técnicas, com diferença significativa na frequência de vimentina, p53 e HER-2, de forma aumentada nos cultivos. A doxorrubicina apresentou alta eficácia nas células tumorais, mas sem diferença entre os tipos histológicos, porém cada cultivo comportou-se distintamente quando analisado em isolado. Conclui-se que é possível cultivar primariamente diferentes tumores mamários caninos utilizando-se meio de cultivo básico; os testes de expressão gênicas são mais indicados para análises comparativas entre tumor e seu respectivo cultivo e, quanto maior a concentração do quimioterápico, menor a viabilidade celularCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Universidade Estadual Paulista (Unesp)Andrade, Alexandre Lima de [UNESP]Silva, Tereza Cristina Cardoso da [UNESP]Universidade Estadual Paulista (Unesp)Lima, Silmara Sanae Sakamoto de [UNESP]2015-10-06T13:03:11Z2015-10-06T13:03:11Z2014-08-09info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesis89 f. : il. + 1 CD-ROMapplication/pdfLIMA, Silmara Sanae Sakamoto de. Tumor mamário canino: estudo in vitro, imunomarcação e ação da doxorrubicina. 2013. 89 f. Tese (doutorado) - Universidade Estadual Paulista Júlio de Mesquita Filho, Faculdade de Medicina Veterinária, 2013.http://hdl.handle.net/11449/128081000850064http://www.athena.biblioteca.unesp.br/exlibris/bd/cathedra/16-09-2015/000850064.pdf33004021075P86404176495306171Alephreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPporinfo:eu-repo/semantics/openAccess2023-11-16T06:11:40Zoai:repositorio.unesp.br:11449/128081Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462023-11-16T06:11:40Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Tumor mamário canino: estudo in vitro, imunomarcação e ação da doxorrubicina
title Tumor mamário canino: estudo in vitro, imunomarcação e ação da doxorrubicina
spellingShingle Tumor mamário canino: estudo in vitro, imunomarcação e ação da doxorrubicina
Lima, Silmara Sanae Sakamoto de [UNESP]
Cão
Mamas - Cancer
Quimioterapia veterinaria
Doxorrubicina
Celulas - Cultura e meios de cultura
Dog diseases
title_short Tumor mamário canino: estudo in vitro, imunomarcação e ação da doxorrubicina
title_full Tumor mamário canino: estudo in vitro, imunomarcação e ação da doxorrubicina
title_fullStr Tumor mamário canino: estudo in vitro, imunomarcação e ação da doxorrubicina
title_full_unstemmed Tumor mamário canino: estudo in vitro, imunomarcação e ação da doxorrubicina
title_sort Tumor mamário canino: estudo in vitro, imunomarcação e ação da doxorrubicina
author Lima, Silmara Sanae Sakamoto de [UNESP]
author_facet Lima, Silmara Sanae Sakamoto de [UNESP]
author_role author
dc.contributor.none.fl_str_mv Andrade, Alexandre Lima de [UNESP]
Silva, Tereza Cristina Cardoso da [UNESP]
Universidade Estadual Paulista (Unesp)
dc.contributor.author.fl_str_mv Lima, Silmara Sanae Sakamoto de [UNESP]
dc.subject.por.fl_str_mv Cão
Mamas - Cancer
Quimioterapia veterinaria
Doxorrubicina
Celulas - Cultura e meios de cultura
Dog diseases
topic Cão
Mamas - Cancer
Quimioterapia veterinaria
Doxorrubicina
Celulas - Cultura e meios de cultura
Dog diseases
description Canine mammary tumors represent a high casuistry in veterinary oncology and few studies with cell culture are being employed, mainly with drug application. The purpose was to establish and standardize the cell culture of canine mammary tumors for in vitro efficacy of doxorubicin and to correlate the immunostaining of vimentin, cytokeratin, p53 and HER-2, in paraffin sections of original tumor and the cell cultures derivated. Benign mixed tumor, complex carcinoma, simple carcinoma and espinocelular carcinoma were cultivated from explants and the cells originated were incubated with doxorubicin in different concentrations (control-0.25-0.50-0.75-1.0-2.0μM). The percentual of viable cells were determined by the Trypan blue dye exclusion test and for immunoexpression, the immunohistochemistry was performed for the primary tumor and immunofluorescence, for the cells. A total of 39 tumor samples were collected and, 40% were cultivated up to fifth passages. A different imunnoexpression was observed between methodologies with higher expression at vimentin, p53 and HER-2 groups. It was observed that doxorubicin presented an elevated efficacy in canine mammary cell culture but without difference among the histological groups and each sample presented an individual responsiveness for the same treatment. So, it can be concluded that it was possible to realize a primary cell culture with a simple culture medium formulation; gene expression profiles in tumor tissue and cell culture derived from those tumors are better designated than immunoexpression tests and higher the concentration of doxorubicin, lower the cell viability
publishDate 2014
dc.date.none.fl_str_mv 2014-08-09
2015-10-06T13:03:11Z
2015-10-06T13:03:11Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv LIMA, Silmara Sanae Sakamoto de. Tumor mamário canino: estudo in vitro, imunomarcação e ação da doxorrubicina. 2013. 89 f. Tese (doutorado) - Universidade Estadual Paulista Júlio de Mesquita Filho, Faculdade de Medicina Veterinária, 2013.
http://hdl.handle.net/11449/128081
000850064
http://www.athena.biblioteca.unesp.br/exlibris/bd/cathedra/16-09-2015/000850064.pdf
33004021075P8
6404176495306171
identifier_str_mv LIMA, Silmara Sanae Sakamoto de. Tumor mamário canino: estudo in vitro, imunomarcação e ação da doxorrubicina. 2013. 89 f. Tese (doutorado) - Universidade Estadual Paulista Júlio de Mesquita Filho, Faculdade de Medicina Veterinária, 2013.
000850064
33004021075P8
6404176495306171
url http://hdl.handle.net/11449/128081
http://www.athena.biblioteca.unesp.br/exlibris/bd/cathedra/16-09-2015/000850064.pdf
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 89 f. : il. + 1 CD-ROM
application/pdf
dc.publisher.none.fl_str_mv Universidade Estadual Paulista (Unesp)
publisher.none.fl_str_mv Universidade Estadual Paulista (Unesp)
dc.source.none.fl_str_mv Aleph
reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
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