Unravelling the role of ACOX3 and MCT2 in prostate cancer
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Tipo de documento: | Dissertação |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10773/25169 |
Resumo: | Prostate cancer has a high incidence rate in men, having a great impact on their life quality, being frequently the cause of death. It is a disease which presents challenges in terms of diagnostics and there is still no permanent and effective treatment. Prostate cancer, contrarily to the majority of other tumours, uses branched chain fatty acids as its main energy source in its initial stages. Recently, our group has reported that a protein involved in the metabolism of glucose, MCT2, is overexpressed in prostate cancer and that its presence at the peroxisomes leads to an increase in the β-oxidation of branched chain fatty acids. The goals of this work were to develop two 22Rv1 stable cell lines, one with the ACOX3 gene knocked out and the other with the MCT2 gene knocked out, using the CRISPR-Cas9 system. Additionally, in order to study the role of ACOX3 in prostate cancer, we created a N-terminal Myc-tagged ACOX3 construct. After several attempts, it was not possible to create the stable cell lines. Regarding the overexpression of ACOX3, no differences were observed in the peroxisome’s dynamics, as well as in the proliferation of 22Rv1 cells. The specific role of peroxisomes, namely through the action of ACOX3 and MCT2, in prostate cancer development still requires further research. The CRISPR-Cas9 system remains an interesting tool for these studies, but a refinement of its protocol is necessary in order to achieve a higher efficiency in the future |
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Unravelling the role of ACOX3 and MCT2 in prostate cancerACOX3MCT2Prostate cancerMetabolismβ-oxidationProstate cancer has a high incidence rate in men, having a great impact on their life quality, being frequently the cause of death. It is a disease which presents challenges in terms of diagnostics and there is still no permanent and effective treatment. Prostate cancer, contrarily to the majority of other tumours, uses branched chain fatty acids as its main energy source in its initial stages. Recently, our group has reported that a protein involved in the metabolism of glucose, MCT2, is overexpressed in prostate cancer and that its presence at the peroxisomes leads to an increase in the β-oxidation of branched chain fatty acids. The goals of this work were to develop two 22Rv1 stable cell lines, one with the ACOX3 gene knocked out and the other with the MCT2 gene knocked out, using the CRISPR-Cas9 system. Additionally, in order to study the role of ACOX3 in prostate cancer, we created a N-terminal Myc-tagged ACOX3 construct. After several attempts, it was not possible to create the stable cell lines. Regarding the overexpression of ACOX3, no differences were observed in the peroxisome’s dynamics, as well as in the proliferation of 22Rv1 cells. The specific role of peroxisomes, namely through the action of ACOX3 and MCT2, in prostate cancer development still requires further research. The CRISPR-Cas9 system remains an interesting tool for these studies, but a refinement of its protocol is necessary in order to achieve a higher efficiency in the futureCancro da próstata é um cancro com elevada incidência em homens, tendo um grande impacto na sua qualidade de vida, sendo frequentemente a causa de morte. É uma doença que a nível de diagnóstico apresenta algumas dificuldades e para a qual ainda não existe um tratamento permanente e eficaz. O cancro da próstata, contrariamente à maioria dos tumores, em fases iniciais usa ácidos gordos ramificados como principal fonte de energia. Recentemente o nosso grupo descobriu que uma proteína envolvida no metabolismo da glucose, MCT2, está sobre expressa no cancro da próstata e que a sua presença nos peroxissomas leva a um aumento da β-oxidação de ácidos gordos ramificados. Os objetivos deste trabalho foram construir duas linhas celulares 22Rv1 estáveis uma com o knockout do gene da ACOX3 e uma outra com o knockout do gene do MCT2, usando o sistema CRISPR-Cas9. Em paralelo, para estudar o papel da ACOX3 no cancro da próstata, pretendeu-se criar um construct da ACOX3 com a adição de um Myc-tag no N-terminal desta proteína. Após várias tentativas não foi possível a criação das linhas estáveis. Relativamente, à sobre expressão da ACOX3, não se verificaram diferenças na dinâmica dos peroxissomas, assim como na proliferação das células 22Rv1. O papel específico dos peroxissomas, nomeadamente através da ação de ACOX3 e MCT2, no desenvolvimento de cancro da próstata necessita de mais estudos. O sistema CRISPR-Cas9 continua a ser uma ferramenta apelativa, sendo necessário um refinamento do seu protocolo de modo a ser uma técnica de maior eficácia no futuro2019-01-21T14:29:50Z2018-12-14T00:00:00Z2018-12-14info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10773/25169TID:202238296engOliveira, Rui Pedro Duarteinfo: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:RCAAP2024-02-22T11:49:02Zoai:ria.ua.pt:10773/25169Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T02:58:34.233682Repositório 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 |
Unravelling the role of ACOX3 and MCT2 in prostate cancer |
title |
Unravelling the role of ACOX3 and MCT2 in prostate cancer |
spellingShingle |
Unravelling the role of ACOX3 and MCT2 in prostate cancer Oliveira, Rui Pedro Duarte ACOX3 MCT2 Prostate cancer Metabolism β-oxidation |
title_short |
Unravelling the role of ACOX3 and MCT2 in prostate cancer |
title_full |
Unravelling the role of ACOX3 and MCT2 in prostate cancer |
title_fullStr |
Unravelling the role of ACOX3 and MCT2 in prostate cancer |
title_full_unstemmed |
Unravelling the role of ACOX3 and MCT2 in prostate cancer |
title_sort |
Unravelling the role of ACOX3 and MCT2 in prostate cancer |
author |
Oliveira, Rui Pedro Duarte |
author_facet |
Oliveira, Rui Pedro Duarte |
author_role |
author |
dc.contributor.author.fl_str_mv |
Oliveira, Rui Pedro Duarte |
dc.subject.por.fl_str_mv |
ACOX3 MCT2 Prostate cancer Metabolism β-oxidation |
topic |
ACOX3 MCT2 Prostate cancer Metabolism β-oxidation |
description |
Prostate cancer has a high incidence rate in men, having a great impact on their life quality, being frequently the cause of death. It is a disease which presents challenges in terms of diagnostics and there is still no permanent and effective treatment. Prostate cancer, contrarily to the majority of other tumours, uses branched chain fatty acids as its main energy source in its initial stages. Recently, our group has reported that a protein involved in the metabolism of glucose, MCT2, is overexpressed in prostate cancer and that its presence at the peroxisomes leads to an increase in the β-oxidation of branched chain fatty acids. The goals of this work were to develop two 22Rv1 stable cell lines, one with the ACOX3 gene knocked out and the other with the MCT2 gene knocked out, using the CRISPR-Cas9 system. Additionally, in order to study the role of ACOX3 in prostate cancer, we created a N-terminal Myc-tagged ACOX3 construct. After several attempts, it was not possible to create the stable cell lines. Regarding the overexpression of ACOX3, no differences were observed in the peroxisome’s dynamics, as well as in the proliferation of 22Rv1 cells. The specific role of peroxisomes, namely through the action of ACOX3 and MCT2, in prostate cancer development still requires further research. The CRISPR-Cas9 system remains an interesting tool for these studies, but a refinement of its protocol is necessary in order to achieve a higher efficiency in the future |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-12-14T00:00:00Z 2018-12-14 2019-01-21T14:29:50Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
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http://hdl.handle.net/10773/25169 TID:202238296 |
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http://hdl.handle.net/10773/25169 |
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TID:202238296 |
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eng |
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eng |
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info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
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reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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