Avaliação da angiogênese em resposta ao tratamento com melatonina no câncer de mama: estudo in vitro e in vivo

Detalhes bibliográficos
Autor(a) principal: Jardim-Perassi, Bruna Victorasso [UNESP]
Data de Publicação: 2014
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://hdl.handle.net/11449/110553
Resumo: Breast cancer has high rates of incidence and mortality, and it is the most common cancer among women. The rapid tumor growth results in hypoxia on tumor microenvironment, leading to a cascade of events that induce angiogenesis and subsequent cancer progression. Thus, the identification of therapeutic agents that can inhibit angiogenesis is essential for the control of tumor progression. Exogenous administration of melatonin, a hormone secreted by the pineal gland, has been shown several oncostatics effects on different types of cancers. The aim of this study was to evaluate the effectiveness of melatonin treatment on angiogenesis in breast cancer, in the in vitro and in vivo studies. In the in vitro study, breast cancer cell lines (MCF-7 and MDA-MB-231) were treated with melatonin under cobalt chloride (CoCl2)-induced hypoxic conditions. Cell viability was measured by MTT assay, the expression of hypoxia-inducible factor 1-alpha (HIF-1α) and vascular endothelial growth factor (VEGF-A) was assessed by real-time PCR and immunocytochemistry. Additionally, other proteins involved in angiogenesis were evaluated by the Protein Array. In the in vivo study, the MDA-MB-231 cells were implanted in athymic nude mice, which were treated with melatonin (40 mg/kg) for 21 days. The tumor was measured weekly and evaluation of angiogenesis was performed by single-photon emission computed tomography (SPECT) with Tc-99m-HYNIC-VEGF-c, which is specific for VEGF receptors (VEGFR2/VEGF3). Moreover, VEGFR2, VEGFR3, von Willebrand factor (vWF) and cell proliferation marker (Ki-67) were evaluated in tumor tissue by immunohistochemistry, and other angiogenic proteins by Protein Array. Results from the in vitro study showed that 1 mM of melatonin under hypoxic conditions (200 μM CoCl2) led to decreased cell viability, protein levels of HIF- 1α and gene and protein expression of VEGF-A in both cell lines (p < 0.05). Among other proteins evaluated, melatonin ...
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spelling Avaliação da angiogênese em resposta ao tratamento com melatonina no câncer de mama: estudo in vitro e in vivoGenéticaCancer - Aspectos geneticosMamas - CancerNeovascularizaçãoMelatoninaCancer - Genetic aspectsBreast cancer has high rates of incidence and mortality, and it is the most common cancer among women. The rapid tumor growth results in hypoxia on tumor microenvironment, leading to a cascade of events that induce angiogenesis and subsequent cancer progression. Thus, the identification of therapeutic agents that can inhibit angiogenesis is essential for the control of tumor progression. Exogenous administration of melatonin, a hormone secreted by the pineal gland, has been shown several oncostatics effects on different types of cancers. The aim of this study was to evaluate the effectiveness of melatonin treatment on angiogenesis in breast cancer, in the in vitro and in vivo studies. In the in vitro study, breast cancer cell lines (MCF-7 and MDA-MB-231) were treated with melatonin under cobalt chloride (CoCl2)-induced hypoxic conditions. Cell viability was measured by MTT assay, the expression of hypoxia-inducible factor 1-alpha (HIF-1α) and vascular endothelial growth factor (VEGF-A) was assessed by real-time PCR and immunocytochemistry. Additionally, other proteins involved in angiogenesis were evaluated by the Protein Array. In the in vivo study, the MDA-MB-231 cells were implanted in athymic nude mice, which were treated with melatonin (40 mg/kg) for 21 days. The tumor was measured weekly and evaluation of angiogenesis was performed by single-photon emission computed tomography (SPECT) with Tc-99m-HYNIC-VEGF-c, which is specific for VEGF receptors (VEGFR2/VEGF3). Moreover, VEGFR2, VEGFR3, von Willebrand factor (vWF) and cell proliferation marker (Ki-67) were evaluated in tumor tissue by immunohistochemistry, and other angiogenic proteins by Protein Array. Results from the in vitro study showed that 1 mM of melatonin under hypoxic conditions (200 μM CoCl2) led to decreased cell viability, protein levels of HIF- 1α and gene and protein expression of VEGF-A in both cell lines (p < 0.05). Among other proteins evaluated, melatonin ...O câncer de mama apresenta altas taxas de incidência e mortalidade, sendo a neoplasia mais comum entre as mulheres. O rápido crescimento do tumor resulta em hipóxia no microambiente tumoral, levando a uma cascata de eventos que induzem a angiogênese e conseqüente progressão do câncer. Assim, a identificação de agentes terapêuticos que possam inibir a angiogênese é essencial para o controle da progressão tumoral. Nesse sentido, tem sido demonstrado que a administração exógena da melatonina, um hormônio naturalmente secretado pela glândula pineal, apresenta diversos efeitos oncostáticos em diferentes tipos de câncer. Assim, o objetivo desse estudo foi avaliar a efetividade do tratamento com melatonina sobre a angiogênese do câncer de mama, em estudos in vitro e in vivo. No estudo in vitro, as linhagens de câncer de mama MCF-7 e MDA-MB-231 foram tratadas com melatonina sob condições de hipóxia mimetizadas pelo Cloreto de Cobalto (CoCl2). A viabilidade celular foi verificada pelo ensaio MTT, a expressão do fator de transcrição induzido por hipóxia (HIF-1α) e do fator de crescimento endotelial vascular (VEGF-A) foi avaliada por PCR em tempo real e imunocitoquímica. Além disso, demais proteínas envolvidas na angiogênese foram avaliadas por Protein Array. No estudo in vivo, as células MDA-MB-231 foram implantadas em camundongos nude atímicos, os quais foram tratados com melatonina (40 mg/kg) por 21 dias. O tumor foi medido semanalmente e a avaliação da angiogênese foi realizada pela técnica de tomografia computadorizada por emissão de fóton único (SPECT), com o radiotraçador Tc- 99m-HYNIC-VEGF-c, agente específico para os receptores de VEGF (VEGFR2/VEGF3). Além disso, as proteínas VEGFR2, VEGFR3, fator de fator de von Willebrand (vWF) e marcador de proliferação celular (Ki-67) foram avaliados no tecido tumoral por imuno-histoquímica, e demais proteínas angiogênicas por Protein Array. O estudo in ...Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Estadual Paulista (Unesp)Zuccari, Debora Aparecida Pires de Campos [UNESP]Universidade Estadual Paulista (Unesp)Jardim-Perassi, Bruna Victorasso [UNESP]2014-11-10T11:09:50Z2014-11-10T11:09:50Z2014-05-26info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesis122 f. : il. color., tabs.application/pdfJARDIM-PERASSI, Bruna Victorasso. Avaliação da angiogênese em resposta ao tratamento com melatonina no câncer de mama: estudo in vitro e in vivo. 2014. 122 f. Tese (doutorado) - Universidade Estadual Paulista Julio de Mesquita Filho, Instituto de Biociências, Letras e Ciências Exatas, 2014.http://hdl.handle.net/11449/110553000790294000790294.pdf33004153023P5Alephreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPporinfo:eu-repo/semantics/openAccess2023-11-29T06:17:36Zoai:repositorio.unesp.br:11449/110553Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T19:05:56.564856Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Avaliação da angiogênese em resposta ao tratamento com melatonina no câncer de mama: estudo in vitro e in vivo
title Avaliação da angiogênese em resposta ao tratamento com melatonina no câncer de mama: estudo in vitro e in vivo
spellingShingle Avaliação da angiogênese em resposta ao tratamento com melatonina no câncer de mama: estudo in vitro e in vivo
Jardim-Perassi, Bruna Victorasso [UNESP]
Genética
Cancer - Aspectos geneticos
Mamas - Cancer
Neovascularização
Melatonina
Cancer - Genetic aspects
title_short Avaliação da angiogênese em resposta ao tratamento com melatonina no câncer de mama: estudo in vitro e in vivo
title_full Avaliação da angiogênese em resposta ao tratamento com melatonina no câncer de mama: estudo in vitro e in vivo
title_fullStr Avaliação da angiogênese em resposta ao tratamento com melatonina no câncer de mama: estudo in vitro e in vivo
title_full_unstemmed Avaliação da angiogênese em resposta ao tratamento com melatonina no câncer de mama: estudo in vitro e in vivo
title_sort Avaliação da angiogênese em resposta ao tratamento com melatonina no câncer de mama: estudo in vitro e in vivo
author Jardim-Perassi, Bruna Victorasso [UNESP]
author_facet Jardim-Perassi, Bruna Victorasso [UNESP]
author_role author
dc.contributor.none.fl_str_mv Zuccari, Debora Aparecida Pires de Campos [UNESP]
Universidade Estadual Paulista (Unesp)
dc.contributor.author.fl_str_mv Jardim-Perassi, Bruna Victorasso [UNESP]
dc.subject.por.fl_str_mv Genética
Cancer - Aspectos geneticos
Mamas - Cancer
Neovascularização
Melatonina
Cancer - Genetic aspects
topic Genética
Cancer - Aspectos geneticos
Mamas - Cancer
Neovascularização
Melatonina
Cancer - Genetic aspects
description Breast cancer has high rates of incidence and mortality, and it is the most common cancer among women. The rapid tumor growth results in hypoxia on tumor microenvironment, leading to a cascade of events that induce angiogenesis and subsequent cancer progression. Thus, the identification of therapeutic agents that can inhibit angiogenesis is essential for the control of tumor progression. Exogenous administration of melatonin, a hormone secreted by the pineal gland, has been shown several oncostatics effects on different types of cancers. The aim of this study was to evaluate the effectiveness of melatonin treatment on angiogenesis in breast cancer, in the in vitro and in vivo studies. In the in vitro study, breast cancer cell lines (MCF-7 and MDA-MB-231) were treated with melatonin under cobalt chloride (CoCl2)-induced hypoxic conditions. Cell viability was measured by MTT assay, the expression of hypoxia-inducible factor 1-alpha (HIF-1α) and vascular endothelial growth factor (VEGF-A) was assessed by real-time PCR and immunocytochemistry. Additionally, other proteins involved in angiogenesis were evaluated by the Protein Array. In the in vivo study, the MDA-MB-231 cells were implanted in athymic nude mice, which were treated with melatonin (40 mg/kg) for 21 days. The tumor was measured weekly and evaluation of angiogenesis was performed by single-photon emission computed tomography (SPECT) with Tc-99m-HYNIC-VEGF-c, which is specific for VEGF receptors (VEGFR2/VEGF3). Moreover, VEGFR2, VEGFR3, von Willebrand factor (vWF) and cell proliferation marker (Ki-67) were evaluated in tumor tissue by immunohistochemistry, and other angiogenic proteins by Protein Array. Results from the in vitro study showed that 1 mM of melatonin under hypoxic conditions (200 μM CoCl2) led to decreased cell viability, protein levels of HIF- 1α and gene and protein expression of VEGF-A in both cell lines (p < 0.05). Among other proteins evaluated, melatonin ...
publishDate 2014
dc.date.none.fl_str_mv 2014-11-10T11:09:50Z
2014-11-10T11:09:50Z
2014-05-26
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 JARDIM-PERASSI, Bruna Victorasso. Avaliação da angiogênese em resposta ao tratamento com melatonina no câncer de mama: estudo in vitro e in vivo. 2014. 122 f. Tese (doutorado) - Universidade Estadual Paulista Julio de Mesquita Filho, Instituto de Biociências, Letras e Ciências Exatas, 2014.
http://hdl.handle.net/11449/110553
000790294
000790294.pdf
33004153023P5
identifier_str_mv JARDIM-PERASSI, Bruna Victorasso. Avaliação da angiogênese em resposta ao tratamento com melatonina no câncer de mama: estudo in vitro e in vivo. 2014. 122 f. Tese (doutorado) - Universidade Estadual Paulista Julio de Mesquita Filho, Instituto de Biociências, Letras e Ciências Exatas, 2014.
000790294
000790294.pdf
33004153023P5
url http://hdl.handle.net/11449/110553
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
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dc.format.none.fl_str_mv 122 f. : il. color., 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
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repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
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