Effect of Regucalcin on the expression of oncogenes and tumor suppressor genes in prostate cell lines

Detalhes bibliográficos
Autor(a) principal: Vaz, Cátia Alexandra Vicente
Data de Publicação: 2010
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10400.6/2292
Resumo: Regucalcin (RGN) is a calcium-binding protein playing an important role in maintenance of intracellular calcium homeostasis. Because of its diminished expression with aging it is also designated Senescence-marker-protein (SMP30). In addition, RGN suppresses cell proliferation, inhibits expression of oncogenes, and increases the expression of tumor suppressor genes in hepatoma cell lines. Very recently, our research group demonstrated that RGN expression is diminished in prostate cancer tissues, what suggests that RGN may have a protective role against carcinogenesis and, consequently, loss of regucalcin expression may contribute to tumor development. The present project aims to characterize RGN expression in prostate tissues and cell-lines and to determine the role of RGN on the expression of oncogenes and tumour suppressor genes in neopasic and non-neoplasic prostate cells. The expression of RGN in rat prostate at different post-natal ages determined by quantitative PCR analysis showed a significant increase at 3M old rats, maintaining their expression in 6M-old animals and diminishing in the following stages. In this report, we confirmed RGN protein expression in human cancer prostate tissues, and cells lines by Imunohistochemistry and Western Blot, respectively. To analyze the effect of RGN on the expression of oncogenes (BCL2, Ha-ras and c-myc) and tumor supressor genes (p53 and RB1), pIRES/RGN expression vectors were constructed and used to transfect LnCAP, PC3, PNT1A and PNT2 cells. Fluorescence microscopy analysis showed successful transfection of PC3 and PNT1A cells, with RGN-GFP protein localization in cell nuclei and cytoplasm. Analysis of transfection experiments in LnCAP and PNT2 cells, and determination of the effect of RGN on the expression of tumor related genes are underway. Nevertheless, RGN localization in cell nuclei, suggests their likely influence regulating expression of oncogenes and tumor suppressor genes in prostate cell lines overexpressing RGN.
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spelling Effect of Regucalcin on the expression of oncogenes and tumor suppressor genes in prostate cell linesInfluência da Regucalcina na expressão de oncogenes e genes supressores de tumor em linhas celulares de próstataGenes supressores de tumoresOncogenes - RegucalcinaCancro da prostata - Regucalcina - Genes supressores de tumorRegucalcin (RGN) is a calcium-binding protein playing an important role in maintenance of intracellular calcium homeostasis. Because of its diminished expression with aging it is also designated Senescence-marker-protein (SMP30). In addition, RGN suppresses cell proliferation, inhibits expression of oncogenes, and increases the expression of tumor suppressor genes in hepatoma cell lines. Very recently, our research group demonstrated that RGN expression is diminished in prostate cancer tissues, what suggests that RGN may have a protective role against carcinogenesis and, consequently, loss of regucalcin expression may contribute to tumor development. The present project aims to characterize RGN expression in prostate tissues and cell-lines and to determine the role of RGN on the expression of oncogenes and tumour suppressor genes in neopasic and non-neoplasic prostate cells. The expression of RGN in rat prostate at different post-natal ages determined by quantitative PCR analysis showed a significant increase at 3M old rats, maintaining their expression in 6M-old animals and diminishing in the following stages. In this report, we confirmed RGN protein expression in human cancer prostate tissues, and cells lines by Imunohistochemistry and Western Blot, respectively. To analyze the effect of RGN on the expression of oncogenes (BCL2, Ha-ras and c-myc) and tumor supressor genes (p53 and RB1), pIRES/RGN expression vectors were constructed and used to transfect LnCAP, PC3, PNT1A and PNT2 cells. Fluorescence microscopy analysis showed successful transfection of PC3 and PNT1A cells, with RGN-GFP protein localization in cell nuclei and cytoplasm. Analysis of transfection experiments in LnCAP and PNT2 cells, and determination of the effect of RGN on the expression of tumor related genes are underway. Nevertheless, RGN localization in cell nuclei, suggests their likely influence regulating expression of oncogenes and tumor suppressor genes in prostate cell lines overexpressing RGN.A Regucalcina (RGN) é uma proteína de ligação ao cálcio desempenhando um papel importante na homeostase intracelular deste ião. Devido á diminuição da sua expressão associada ao envelhecimento é também designada como Proteína marcadora de senescência (SMP30). A RGN tem sido associada a funções tais como, supressão da proliferação celular, inibição da expressão de oncogenes e aumento na expressão de genes supressores de tumor em linhas celulares de fígado. Recentemente, o nosso grupo de investigação demonstrou que a expressão da RGN estava diminuida em tecidos de adenocarcinomas de próstata, o que sugere um efeito protector da RGN na carcinogénese, e consequentemente a perda da sua expressão pode estar associada ao desenvolvimento tumoral. O presente trabalho teve como objectivos a caracterização da expressão da RGN em linhas celulares e tecidos de próstata e determinar o papel da RGN na expressão de oncogenes e genes supressores de tumor em linhas celulares da próstata neoplásicas e não-neoplásicas. A expressão da RGN na próstata de rato ao longo do desenvolvimento pós-natal foi estudada por PCR quantitativo. A expressão da RGN aumentou aos 3M de idade, a sua expressão manteve-se elevada até aos 6M e diminuiu nas idades seguintes. Neste estudo foi também possível confirmar a expressão da RGN em tecidos de adenocarcinoma de próstata e em linhas celulares por imunohistoquimica e Western Blot, respectivamente. Para analisar o efeito da RGN na expressão de oncogenes e genes supressores de tumor, foram construidos vectores de expressão pIRES/RGN os quais foram usados para transfectar as linhas celulares LnCAP, PC3, PNT1A e PNT2. A análise efectuada por microscopia de fluorescência demonstrou a localização da proteína RGN-GFP no núcleo e citoplama das células PC3 e PNT1A transfectadas com sucesso. A análise da transfecção das celúlas LnCAP e PNT2, bem como a determinação do efeito da RGN na expressão de genes associados ao desenvolvimento tumoral encontra-se em curso. No entanto, os resultados que demonstram a localização da RGN no núcleo, sugerem o seu possível envolvimento na regulação da expressão de oncogenes e genes supressores de tumor em linhas celulares da próstata que sobre-expressam a RGN.Socorro, Sílvia Cristina da Cruz MarquesBaptista, Cláudio Jorge MaiauBibliorumVaz, Cátia Alexandra Vicente2014-09-17T12:25:37Z201020102010-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10400.6/2292enginfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-12-15T09:38:15Zoai:ubibliorum.ubi.pt:10400.6/2292Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T00:43:59.499806Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Effect of Regucalcin on the expression of oncogenes and tumor suppressor genes in prostate cell lines
Influência da Regucalcina na expressão de oncogenes e genes supressores de tumor em linhas celulares de próstata
title Effect of Regucalcin on the expression of oncogenes and tumor suppressor genes in prostate cell lines
spellingShingle Effect of Regucalcin on the expression of oncogenes and tumor suppressor genes in prostate cell lines
Vaz, Cátia Alexandra Vicente
Genes supressores de tumores
Oncogenes - Regucalcina
Cancro da prostata - Regucalcina - Genes supressores de tumor
title_short Effect of Regucalcin on the expression of oncogenes and tumor suppressor genes in prostate cell lines
title_full Effect of Regucalcin on the expression of oncogenes and tumor suppressor genes in prostate cell lines
title_fullStr Effect of Regucalcin on the expression of oncogenes and tumor suppressor genes in prostate cell lines
title_full_unstemmed Effect of Regucalcin on the expression of oncogenes and tumor suppressor genes in prostate cell lines
title_sort Effect of Regucalcin on the expression of oncogenes and tumor suppressor genes in prostate cell lines
author Vaz, Cátia Alexandra Vicente
author_facet Vaz, Cátia Alexandra Vicente
author_role author
dc.contributor.none.fl_str_mv Socorro, Sílvia Cristina da Cruz Marques
Baptista, Cláudio Jorge Maia
uBibliorum
dc.contributor.author.fl_str_mv Vaz, Cátia Alexandra Vicente
dc.subject.por.fl_str_mv Genes supressores de tumores
Oncogenes - Regucalcina
Cancro da prostata - Regucalcina - Genes supressores de tumor
topic Genes supressores de tumores
Oncogenes - Regucalcina
Cancro da prostata - Regucalcina - Genes supressores de tumor
description Regucalcin (RGN) is a calcium-binding protein playing an important role in maintenance of intracellular calcium homeostasis. Because of its diminished expression with aging it is also designated Senescence-marker-protein (SMP30). In addition, RGN suppresses cell proliferation, inhibits expression of oncogenes, and increases the expression of tumor suppressor genes in hepatoma cell lines. Very recently, our research group demonstrated that RGN expression is diminished in prostate cancer tissues, what suggests that RGN may have a protective role against carcinogenesis and, consequently, loss of regucalcin expression may contribute to tumor development. The present project aims to characterize RGN expression in prostate tissues and cell-lines and to determine the role of RGN on the expression of oncogenes and tumour suppressor genes in neopasic and non-neoplasic prostate cells. The expression of RGN in rat prostate at different post-natal ages determined by quantitative PCR analysis showed a significant increase at 3M old rats, maintaining their expression in 6M-old animals and diminishing in the following stages. In this report, we confirmed RGN protein expression in human cancer prostate tissues, and cells lines by Imunohistochemistry and Western Blot, respectively. To analyze the effect of RGN on the expression of oncogenes (BCL2, Ha-ras and c-myc) and tumor supressor genes (p53 and RB1), pIRES/RGN expression vectors were constructed and used to transfect LnCAP, PC3, PNT1A and PNT2 cells. Fluorescence microscopy analysis showed successful transfection of PC3 and PNT1A cells, with RGN-GFP protein localization in cell nuclei and cytoplasm. Analysis of transfection experiments in LnCAP and PNT2 cells, and determination of the effect of RGN on the expression of tumor related genes are underway. Nevertheless, RGN localization in cell nuclei, suggests their likely influence regulating expression of oncogenes and tumor suppressor genes in prostate cell lines overexpressing RGN.
publishDate 2010
dc.date.none.fl_str_mv 2010
2010
2010-01-01T00:00:00Z
2014-09-17T12:25:37Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
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status_str publishedVersion
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url http://hdl.handle.net/10400.6/2292
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
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