Expressão de Indoleamina 2,3 Dioxigenase (IDO) em células T24 de carcinoma de bexiga urinária submetidas à hipóxia: correlação com Fator de Crescimento Endotelial Vascular (VEGF)
Autor(a) principal: | |
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Data de Publicação: | 2015 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Biblioteca Digital de Teses e Dissertações da Uninove |
Texto Completo: | http://bibliotecatede.uninove.br/handle/tede/3315 |
Resumo: | The urinary bladder cancer is the most common malignancy of the urinary system and for his growth and progression, angiogenesis is necessary. A indoleamine 2,3-dioxygenase (IDO), a molecule known for its immunomodulatory properties arises as a potential modulator of angiogenesis. Its expression is associated with pro-angiogenic factors in different types of cancer, but in bladder cancer has not yet been explored. The aim of this study was to analyze IDO expression in T24 cells of bladder carcinoma in situation of hypoxia in order to correlate it with angiogenesis-inducing factors. T24 cells were subjected to hypoxia by AnaeroGen product during four periods of incubation (1 h, 8 h, 24 h and 48 h). Real time RT-PCR was performed to analyze the expression of VEGF, HIF and IDO. Cells without hypoxia were used as controls. As a second step, the cells were treated with the IDO inhibitor (1-methyl tryptophan) and subjected to hypoxia or not for the evaluation of VEGF expression. In addition, HUVEC cells were treated with the IDO inhibitor to assess cell viability. In "in vivo" step, Balb/c nude received T24 cells under the kidney capsule for the establishment of a model that would allow assessment of tumor neovasculature (30 days). The characterization of neovascularization was performed by immunohistochemistry. The expression of VEGF was significantly increased over the incubation periods, and time-dependent. But the expression of HIF and IDO did not change significantly. However, there was a positive correlation between HIF and IDO (correlation coefficient 0.615, Spearman). Treatment with the IDO inhibitor significantly decreased VEGF expression in T24 cells subjected to hypoxia, in addition to reducing the viability of HUVEC cells. Renal subcapsular inoculation of T24 cells is an interesting model for the study of the formation of new vessels, but animals treated with IDO inhibitor did not shown formation of subcapsular tumor, only infiltrates that not allowed the vascular quantification. The study concludes that IDO is correlated with factors that induce angiogenesis in hypoxic condition and its inhibition decreases the expression of VEGF and viability of endothelial cells. Inhibition of IDO may be a promising alternative for inhibiting angiogenesis in bladder tumors. |
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Dellê, Humbertohttp://lattes.cnpq.br/7435662740477057Vidsiunas, Alex Korshttp://lattes.cnpq.br/4171000415247040Dellê, Humbertohttp://lattes.cnpq.br/7435662740477057Reis, Sabrina Thalita doshttp://lattes.cnpq.br/7256172784284719Zamuner, Stella Reginahttp://lattes.cnpq.br/1935805744318404http://lattes.cnpq.br/9267132013785790Cesário, Jonas Magno dos Santos2024-04-08T21:05:06Z2015-02-03Cesário, Jonas Magno dos Santos. Expressão de Indoleamina 2,3 Dioxigenase (IDO) em células T24 de carcinoma de bexiga urinária submetidas à hipóxia: correlação com Fator de Crescimento Endotelial Vascular (VEGF). 2015. 51 f. Dissertação( Programa de Mestrado em Medicina) - Universidade Nove de Julho, São Paulo.http://bibliotecatede.uninove.br/handle/tede/3315The urinary bladder cancer is the most common malignancy of the urinary system and for his growth and progression, angiogenesis is necessary. A indoleamine 2,3-dioxygenase (IDO), a molecule known for its immunomodulatory properties arises as a potential modulator of angiogenesis. Its expression is associated with pro-angiogenic factors in different types of cancer, but in bladder cancer has not yet been explored. The aim of this study was to analyze IDO expression in T24 cells of bladder carcinoma in situation of hypoxia in order to correlate it with angiogenesis-inducing factors. T24 cells were subjected to hypoxia by AnaeroGen product during four periods of incubation (1 h, 8 h, 24 h and 48 h). Real time RT-PCR was performed to analyze the expression of VEGF, HIF and IDO. Cells without hypoxia were used as controls. As a second step, the cells were treated with the IDO inhibitor (1-methyl tryptophan) and subjected to hypoxia or not for the evaluation of VEGF expression. In addition, HUVEC cells were treated with the IDO inhibitor to assess cell viability. In "in vivo" step, Balb/c nude received T24 cells under the kidney capsule for the establishment of a model that would allow assessment of tumor neovasculature (30 days). The characterization of neovascularization was performed by immunohistochemistry. The expression of VEGF was significantly increased over the incubation periods, and time-dependent. But the expression of HIF and IDO did not change significantly. However, there was a positive correlation between HIF and IDO (correlation coefficient 0.615, Spearman). Treatment with the IDO inhibitor significantly decreased VEGF expression in T24 cells subjected to hypoxia, in addition to reducing the viability of HUVEC cells. Renal subcapsular inoculation of T24 cells is an interesting model for the study of the formation of new vessels, but animals treated with IDO inhibitor did not shown formation of subcapsular tumor, only infiltrates that not allowed the vascular quantification. The study concludes that IDO is correlated with factors that induce angiogenesis in hypoxic condition and its inhibition decreases the expression of VEGF and viability of endothelial cells. Inhibition of IDO may be a promising alternative for inhibiting angiogenesis in bladder tumors.O câncer de bexiga urinária é a neoplasia maligna mais comum do sistema urinário e para seu crescimento e progressão, a angiogênese é necessária. A Indoleamina 2,3-dioxigenase (IDO), uma molécula reconhecida por suas propriedades imunomoduladoras e surge como um potencial modulador da angiogênese. Sua expressão está associada com fatores pró-angiogênicos em tipos diferentes de câncer, porém no câncer de bexiga não foi ainda explorada. O objetivo do presente estudo foi analisar a expressão de IDO em células T24 de carcinoma de bexiga urinária em situação de hipóxia, a fim de correlacioná-la com fatores de indução de angiogênese. Células T24 foram submetidas à hipóxia através do produto AnaeroGen durante quatro períodos de incubação (1 h, 8 h, 24 h e 48 h). RT-PCR em tempo real foi realizada para análise da expressão de VEGF, HIF e IDO. Células sem hipóxia foram usadas como controle. Como segunda etapa, as células foram tratadas com o inibidor da IDO (1-metil triptofano) e submetidas ou não à hipóxia para a avaliação da expressão do VEGF. Adicionalmente, células HUVEC foram tratadas com o inibidor da IDO para avaliação da viabilidade celular. Na fase “in vivo”, camundongos Balb/c nude receberam células T24 sob a cápsula renal para o estabelecimento de um modelo que permitisse a avaliação de neovasos tumorais (30 dias). A caracterização dos neovasos foi feita por imuno-histoquímica. A expressão de VEGF aumentou significativamente ao longo dos períodos de incubação, sendo tempo-dependente. Já a expressão de HIF e IDO não sofreu alteração significativa. Entretanto, houve uma correlação positiva entre HIF e IDO (coeficiente de correlação 0,615, Spearman). O tratamento com inibidor da IDO diminuiu significativamente a expressão de VEGF em células T24 submetidas à hipóxia, além de diminuir a viabilidade de células HUVEC. A inoculação subcapsular renal de células T24 é um modelo interessante para o estudo da formação de novos vasos, porém animais tratados com inibidor da IDO não apresentaram formação de tumor subcapsular, apenas infiltrados que não permitiram a quantificação vascular. O estudo conclui que a IDO correlaciona-se com fatores indutores de angiogênese na situação de hipóxia e sua inibição diminui a expressão de VEGF e a viabilidade de células endoteliais. A inibição da IDO pode ser uma alternativa promissora para inibir angiogênese em tumores de bexiga urinária.Submitted by Nadir Basilio (nadirsb@uninove.br) on 2024-04-08T21:05:06Z No. of bitstreams: 1 Jonas Magno dos Santos Cesário.pdf: 1263356 bytes, checksum: 026c1d1514287bef3c546b3c4e43dd13 (MD5)Made available in DSpace on 2024-04-08T21:05:06Z (GMT). No. of bitstreams: 1 Jonas Magno dos Santos Cesário.pdf: 1263356 bytes, checksum: 026c1d1514287bef3c546b3c4e43dd13 (MD5) Previous issue date: 2015-02-03application/pdfporUniversidade Nove de JulhoPrograma de Mestrado em MedicinaUNINOVEBrasilSaúdecâncer de bexiga urináriaangiogênesehipóxiaindoleamina 2,3-dioxigenaseVEGFbladder cancerangiogenesishypoxiaindoleamine 2,3-dioxygenaseVEGFCIENCIAS DA SAUDEExpressão de Indoleamina 2,3 Dioxigenase (IDO) em células T24 de carcinoma de bexiga urinária submetidas à hipóxia: correlação com Fator de Crescimento Endotelial Vascular (VEGF)Indoleamine 2,3 Dioxygenase (IDO) expression in T24 urinary bladder carcinoma cells submitted to hypoxia: correlation with Vascular Endothelial Growth Factor (VEGF)info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesis8765449414823306929600info:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações da Uninoveinstname:Universidade Nove de Julho (UNINOVE)instacron:UNINOVEORIGINALJonas Magno dos Santos Cesário.pdfJonas Magno dos Santos Cesário.pdfapplication/pdf1263356http://localhost:8080/tede/bitstream/tede/3315/2/Jonas+Magno+dos+Santos+Ces%C3%A1rio.pdf026c1d1514287bef3c546b3c4e43dd13MD52LICENSElicense.txtlicense.txttext/plain; charset=utf-82165http://localhost:8080/tede/bitstream/tede/3315/1/license.txtbd3efa91386c1718a7f26a329fdcb468MD51tede/33152024-04-08 18:05:06.778oai:localhost: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Biblioteca Digital de Teses e Dissertaçõeshttp://bibliotecatede.uninove.br/PRIhttp://bibliotecatede.uninove.br/oai/requestbibliotecatede@uninove.br||bibliotecatede@uninove.bropendoar:2024-04-08T21:05:06Biblioteca Digital de Teses e Dissertações da Uninove - Universidade Nove de Julho (UNINOVE)false |
dc.title.por.fl_str_mv |
Expressão de Indoleamina 2,3 Dioxigenase (IDO) em células T24 de carcinoma de bexiga urinária submetidas à hipóxia: correlação com Fator de Crescimento Endotelial Vascular (VEGF) |
dc.title.alternative.eng.fl_str_mv |
Indoleamine 2,3 Dioxygenase (IDO) expression in T24 urinary bladder carcinoma cells submitted to hypoxia: correlation with Vascular Endothelial Growth Factor (VEGF) |
title |
Expressão de Indoleamina 2,3 Dioxigenase (IDO) em células T24 de carcinoma de bexiga urinária submetidas à hipóxia: correlação com Fator de Crescimento Endotelial Vascular (VEGF) |
spellingShingle |
Expressão de Indoleamina 2,3 Dioxigenase (IDO) em células T24 de carcinoma de bexiga urinária submetidas à hipóxia: correlação com Fator de Crescimento Endotelial Vascular (VEGF) Cesário, Jonas Magno dos Santos câncer de bexiga urinária angiogênese hipóxia indoleamina 2,3-dioxigenase VEGF bladder cancer angiogenesis hypoxia indoleamine 2,3-dioxygenase VEGF CIENCIAS DA SAUDE |
title_short |
Expressão de Indoleamina 2,3 Dioxigenase (IDO) em células T24 de carcinoma de bexiga urinária submetidas à hipóxia: correlação com Fator de Crescimento Endotelial Vascular (VEGF) |
title_full |
Expressão de Indoleamina 2,3 Dioxigenase (IDO) em células T24 de carcinoma de bexiga urinária submetidas à hipóxia: correlação com Fator de Crescimento Endotelial Vascular (VEGF) |
title_fullStr |
Expressão de Indoleamina 2,3 Dioxigenase (IDO) em células T24 de carcinoma de bexiga urinária submetidas à hipóxia: correlação com Fator de Crescimento Endotelial Vascular (VEGF) |
title_full_unstemmed |
Expressão de Indoleamina 2,3 Dioxigenase (IDO) em células T24 de carcinoma de bexiga urinária submetidas à hipóxia: correlação com Fator de Crescimento Endotelial Vascular (VEGF) |
title_sort |
Expressão de Indoleamina 2,3 Dioxigenase (IDO) em células T24 de carcinoma de bexiga urinária submetidas à hipóxia: correlação com Fator de Crescimento Endotelial Vascular (VEGF) |
author |
Cesário, Jonas Magno dos Santos |
author_facet |
Cesário, Jonas Magno dos Santos |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Dellê, Humberto |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/7435662740477057 |
dc.contributor.advisor-co1.fl_str_mv |
Vidsiunas, Alex Kors |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://lattes.cnpq.br/4171000415247040 |
dc.contributor.referee1.fl_str_mv |
Dellê, Humberto |
dc.contributor.referee1Lattes.fl_str_mv |
http://lattes.cnpq.br/7435662740477057 |
dc.contributor.referee2.fl_str_mv |
Reis, Sabrina Thalita dos |
dc.contributor.referee2Lattes.fl_str_mv |
http://lattes.cnpq.br/7256172784284719 |
dc.contributor.referee3.fl_str_mv |
Zamuner, Stella Regina |
dc.contributor.referee3Lattes.fl_str_mv |
http://lattes.cnpq.br/1935805744318404 |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/9267132013785790 |
dc.contributor.author.fl_str_mv |
Cesário, Jonas Magno dos Santos |
contributor_str_mv |
Dellê, Humberto Vidsiunas, Alex Kors Dellê, Humberto Reis, Sabrina Thalita dos Zamuner, Stella Regina |
dc.subject.por.fl_str_mv |
câncer de bexiga urinária angiogênese hipóxia indoleamina 2,3-dioxigenase VEGF |
topic |
câncer de bexiga urinária angiogênese hipóxia indoleamina 2,3-dioxigenase VEGF bladder cancer angiogenesis hypoxia indoleamine 2,3-dioxygenase VEGF CIENCIAS DA SAUDE |
dc.subject.eng.fl_str_mv |
bladder cancer angiogenesis hypoxia indoleamine 2,3-dioxygenase VEGF |
dc.subject.cnpq.fl_str_mv |
CIENCIAS DA SAUDE |
description |
The urinary bladder cancer is the most common malignancy of the urinary system and for his growth and progression, angiogenesis is necessary. A indoleamine 2,3-dioxygenase (IDO), a molecule known for its immunomodulatory properties arises as a potential modulator of angiogenesis. Its expression is associated with pro-angiogenic factors in different types of cancer, but in bladder cancer has not yet been explored. The aim of this study was to analyze IDO expression in T24 cells of bladder carcinoma in situation of hypoxia in order to correlate it with angiogenesis-inducing factors. T24 cells were subjected to hypoxia by AnaeroGen product during four periods of incubation (1 h, 8 h, 24 h and 48 h). Real time RT-PCR was performed to analyze the expression of VEGF, HIF and IDO. Cells without hypoxia were used as controls. As a second step, the cells were treated with the IDO inhibitor (1-methyl tryptophan) and subjected to hypoxia or not for the evaluation of VEGF expression. In addition, HUVEC cells were treated with the IDO inhibitor to assess cell viability. In "in vivo" step, Balb/c nude received T24 cells under the kidney capsule for the establishment of a model that would allow assessment of tumor neovasculature (30 days). The characterization of neovascularization was performed by immunohistochemistry. The expression of VEGF was significantly increased over the incubation periods, and time-dependent. But the expression of HIF and IDO did not change significantly. However, there was a positive correlation between HIF and IDO (correlation coefficient 0.615, Spearman). Treatment with the IDO inhibitor significantly decreased VEGF expression in T24 cells subjected to hypoxia, in addition to reducing the viability of HUVEC cells. Renal subcapsular inoculation of T24 cells is an interesting model for the study of the formation of new vessels, but animals treated with IDO inhibitor did not shown formation of subcapsular tumor, only infiltrates that not allowed the vascular quantification. The study concludes that IDO is correlated with factors that induce angiogenesis in hypoxic condition and its inhibition decreases the expression of VEGF and viability of endothelial cells. Inhibition of IDO may be a promising alternative for inhibiting angiogenesis in bladder tumors. |
publishDate |
2015 |
dc.date.issued.fl_str_mv |
2015-02-03 |
dc.date.accessioned.fl_str_mv |
2024-04-08T21:05:06Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/masterThesis |
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dc.identifier.citation.fl_str_mv |
Cesário, Jonas Magno dos Santos. Expressão de Indoleamina 2,3 Dioxigenase (IDO) em células T24 de carcinoma de bexiga urinária submetidas à hipóxia: correlação com Fator de Crescimento Endotelial Vascular (VEGF). 2015. 51 f. Dissertação( Programa de Mestrado em Medicina) - Universidade Nove de Julho, São Paulo. |
dc.identifier.uri.fl_str_mv |
http://bibliotecatede.uninove.br/handle/tede/3315 |
identifier_str_mv |
Cesário, Jonas Magno dos Santos. Expressão de Indoleamina 2,3 Dioxigenase (IDO) em células T24 de carcinoma de bexiga urinária submetidas à hipóxia: correlação com Fator de Crescimento Endotelial Vascular (VEGF). 2015. 51 f. Dissertação( Programa de Mestrado em Medicina) - Universidade Nove de Julho, São Paulo. |
url |
http://bibliotecatede.uninove.br/handle/tede/3315 |
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Saúde |
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