Análise funcional do gene GPC3 em carcinoma renal de células claras

Detalhes bibliográficos
Ano de defesa: 2013
Autor(a) principal: Valsechi, Marina Curado [UNESP]
Orientador(a): Não Informado pela instituição
Banca de defesa: Não Informado pela instituição
Tipo de documento: Tese
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Estadual Paulista (Unesp)
Programa de Pós-Graduação: Não Informado pela instituição
Departamento: Não Informado pela instituição
País: Não Informado pela instituição
Palavras-chave em Português:
Cancer
Rins
Carcinoma de células renais
Expressão gênica
Regulamento genetico
Link de acesso: http://hdl.handle.net/11449/127546
Resumo: Background: GPC3 (Glypican 3) is a member of the family of glypican heparan sulfate proteoglycans (HSPG). The GPC3 gene may play a role in controlling cell migration, negative regulation of cell growth and induction of apoptosis. This gene is reported to be downregulated in several cancers, which can result in uncontrolled cell growth and which can also contribute to the malignant phenotype of some tumors. The purpose of this study was to analyze the mechanism of action of the GPC3 gene in clear cell renal cell carcinoma (CCRCC). Methods: First, we constructed the expression vector and transfected renal carcinoma cell lines. GPC3 expression was analyzed using qRT-PCR and immunohistochemistry. We evaluated cell proliferation using MTT and colony formation assays. Apoptosis and cell cycle analyses were evaluated using flow cytometry. Results: We observed that the GPC3 gene was downexpressed in the clear cell renal cell carcinoma samples and in cell lines, which were both compared to normal renal samples. We observed that GPC3 mRNA expression and protein levels increased after the transfection into ACHN and 786-O cell lines. We found that the cell proliferation rate decreased in cells overexpressing GPC3 in both cell lines, ACHN and 786-O (p < 0.01). Also, apoptosis in the renal cell carcinoma cell line was not observed in cells overexpressing GPC3 (p > 0.05), but there was an increase in the cell population during the G1 phase in the cell cycle (p< 0.05). Conclusion: We suggest that the GPC3 gene reduced the cell proliferation rate through cell cycle arrest during the G1 phase in renal cell carcinoma
id UNSP_5fd92635d4ab5fc1784ee8290325fd73
oai_identifier_str oai:repositorio.unesp.br:11449/127546
network_acronym_str UNSP
network_name_str Repositório Institucional da UNESP
repository_id_str
spelling Análise funcional do gene GPC3 em carcinoma renal de células clarasCancerRinsCarcinoma de células renaisExpressão gênicaRegulamento geneticoBackground: GPC3 (Glypican 3) is a member of the family of glypican heparan sulfate proteoglycans (HSPG). The GPC3 gene may play a role in controlling cell migration, negative regulation of cell growth and induction of apoptosis. This gene is reported to be downregulated in several cancers, which can result in uncontrolled cell growth and which can also contribute to the malignant phenotype of some tumors. The purpose of this study was to analyze the mechanism of action of the GPC3 gene in clear cell renal cell carcinoma (CCRCC). Methods: First, we constructed the expression vector and transfected renal carcinoma cell lines. GPC3 expression was analyzed using qRT-PCR and immunohistochemistry. We evaluated cell proliferation using MTT and colony formation assays. Apoptosis and cell cycle analyses were evaluated using flow cytometry. Results: We observed that the GPC3 gene was downexpressed in the clear cell renal cell carcinoma samples and in cell lines, which were both compared to normal renal samples. We observed that GPC3 mRNA expression and protein levels increased after the transfection into ACHN and 786-O cell lines. We found that the cell proliferation rate decreased in cells overexpressing GPC3 in both cell lines, ACHN and 786-O (p < 0.01). Also, apoptosis in the renal cell carcinoma cell line was not observed in cells overexpressing GPC3 (p > 0.05), but there was an increase in the cell population during the G1 phase in the cell cycle (p< 0.05). Conclusion: We suggest that the GPC3 gene reduced the cell proliferation rate through cell cycle arrest during the G1 phase in renal cell carcinomaGPC3 (Glipican-3) é membro de uma família de proteoglicano de heparina sulfatada (HSPG). O GPC3 pode atuar controlando a migração celular, regulação negativa do crescimento celular e indução de apoptose. Esse gene é relatado por estar hipoexpresso em vários tipos de cânceres, o que pode resultar no crescimento celular descontrolado e contribuir para o fenótipo maligno de alguns tumores. O objetivo desse estudo foi analisar o mecanismo de ação do gene GPC3 em carcinoma renal de células claras (CCRCC). Primeiramente, foi construído o vetor de expressão e transfectado nas linhagens celulares de carcinoma renal ACHN e 786-O. A expressão de GPC3 foi analisada usando qRT-PCR e imunohistoquímica. A proliferação celular foi avaliada usando o MTT e o ensaio de formação de colônia. Análises de apoptose e ciclo celular foram avaliadas por citometria de fluxo. Foi observado que o gene GPC3 estava com baixa expressão em amostras de carcinoma renal de células claras e nas linhagens celulares quando comparado com amostras renais normais. Foi observado que a expressão de RNAm e os níveis de proteína GPC3 aumentaram após a transfecção com o vetor de expressão contendo a ORF de GPC3 nas linhagens celulares. A taxa de proliferação celular diminuiu nas células superexpressando GPC3 em ambas as linhagens, ACHN e 786-O (p<0,01). A apoptose não foi observada nas linhagens celulares de carcinoma renal superexpressando GPC3 (p > 0,05); entretanto, ocorreu um aumento na população de células na fase G1 do ciclo celular (p< 0,05). Esses resultados sugerem que o gene GPC3 reduz a taxa de proliferação celular por meio da parada do ciclo celular na fase G1 em carcinoma renalFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Estadual Paulista (Unesp)Rahal, Paula [UNESP]Universidade Estadual Paulista (Unesp)Valsechi, Marina Curado [UNESP]2015-09-17T15:24:08Z2015-09-17T15:24:08Z2013-08-14info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesis102 f. : il. grafs. tabs.application/pdfVALSECHI, Marina Curado. Análise funcional do gene GPC3 em carcinoma renal de células claras. 2013. 102 f. Tese (doutorado) - Universidade Estadual Paulista Julio de Mesquita Filho, Instituto de Biociências, Letras e Ciências Exatas, 2013.http://hdl.handle.net/11449/127546000846657000846657.pdf33004153023P579910823626712120000-0001-5693-6148Alephreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPporinfo:eu-repo/semantics/openAccess2024-11-05T17:49:26Zoai:repositorio.unesp.br:11449/127546Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestrepositoriounesp@unesp.bropendoar:29462024-11-05T17:49:26Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Análise funcional do gene GPC3 em carcinoma renal de células claras
title Análise funcional do gene GPC3 em carcinoma renal de células claras
spellingShingle Análise funcional do gene GPC3 em carcinoma renal de células claras
Valsechi, Marina Curado [UNESP]
Cancer
Rins
Carcinoma de células renais
Expressão gênica
Regulamento genetico
title_short Análise funcional do gene GPC3 em carcinoma renal de células claras
title_full Análise funcional do gene GPC3 em carcinoma renal de células claras
title_fullStr Análise funcional do gene GPC3 em carcinoma renal de células claras
title_full_unstemmed Análise funcional do gene GPC3 em carcinoma renal de células claras
title_sort Análise funcional do gene GPC3 em carcinoma renal de células claras
author Valsechi, Marina Curado [UNESP]
author_facet Valsechi, Marina Curado [UNESP]
author_role author
dc.contributor.none.fl_str_mv Rahal, Paula [UNESP]
Universidade Estadual Paulista (Unesp)
dc.contributor.author.fl_str_mv Valsechi, Marina Curado [UNESP]
dc.subject.por.fl_str_mv Cancer
Rins
Carcinoma de células renais
Expressão gênica
Regulamento genetico
topic Cancer
Rins
Carcinoma de células renais
Expressão gênica
Regulamento genetico
description Background: GPC3 (Glypican 3) is a member of the family of glypican heparan sulfate proteoglycans (HSPG). The GPC3 gene may play a role in controlling cell migration, negative regulation of cell growth and induction of apoptosis. This gene is reported to be downregulated in several cancers, which can result in uncontrolled cell growth and which can also contribute to the malignant phenotype of some tumors. The purpose of this study was to analyze the mechanism of action of the GPC3 gene in clear cell renal cell carcinoma (CCRCC). Methods: First, we constructed the expression vector and transfected renal carcinoma cell lines. GPC3 expression was analyzed using qRT-PCR and immunohistochemistry. We evaluated cell proliferation using MTT and colony formation assays. Apoptosis and cell cycle analyses were evaluated using flow cytometry. Results: We observed that the GPC3 gene was downexpressed in the clear cell renal cell carcinoma samples and in cell lines, which were both compared to normal renal samples. We observed that GPC3 mRNA expression and protein levels increased after the transfection into ACHN and 786-O cell lines. We found that the cell proliferation rate decreased in cells overexpressing GPC3 in both cell lines, ACHN and 786-O (p < 0.01). Also, apoptosis in the renal cell carcinoma cell line was not observed in cells overexpressing GPC3 (p > 0.05), but there was an increase in the cell population during the G1 phase in the cell cycle (p< 0.05). Conclusion: We suggest that the GPC3 gene reduced the cell proliferation rate through cell cycle arrest during the G1 phase in renal cell carcinoma
publishDate 2013
dc.date.none.fl_str_mv 2013-08-14
2015-09-17T15:24:08Z
2015-09-17T15:24:08Z
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 VALSECHI, Marina Curado. Análise funcional do gene GPC3 em carcinoma renal de células claras. 2013. 102 f. Tese (doutorado) - Universidade Estadual Paulista Julio de Mesquita Filho, Instituto de Biociências, Letras e Ciências Exatas, 2013.
http://hdl.handle.net/11449/127546
000846657
000846657.pdf
33004153023P5
7991082362671212
0000-0001-5693-6148
identifier_str_mv VALSECHI, Marina Curado. Análise funcional do gene GPC3 em carcinoma renal de células claras. 2013. 102 f. Tese (doutorado) - Universidade Estadual Paulista Julio de Mesquita Filho, Instituto de Biociências, Letras e Ciências Exatas, 2013.
000846657
000846657.pdf
33004153023P5
7991082362671212
0000-0001-5693-6148
url http://hdl.handle.net/11449/127546
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 102 f. : il. grafs. tabs.
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)
repository.mail.fl_str_mv repositoriounesp@unesp.br
_version_ 1854954516624965632

Registros relacionados