NRF2 and Keap1 Genetic Polymorphisms in Breast Cancer
Autor(a) principal: | |
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Data de Publicação: | 2022 |
Tipo de documento: | Dissertação |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10400.6/12896 |
Resumo: | Cancer is one of the diseases with the highest mortality rate, with breast cancer being the most frequent and with the highest mortality rate in females. Human exposure to oxidative stress may be correlated with the development of several pathologies, including cancer. The pathways involved in oxidative stress represent a risk factor for the development of breast cancer. One of the most important factors in oxidative stress regulation is the NF-E2-related factor 2 (Nrf2)-Kelch-like ECH-associated protein 1 (Keap1) (Nrf2-Keap1) complex. The regulation of this complex may be compromised, due to somatic mutations that influence the loss of negative regulation or the presence of Single Nucleotide Polymorphisms in the promoter region of the NRF2 gene, which may be related to changes in the expression of this transcription factor and its biological consequences. Genetic polymorphisms have become potential research targets in breast cancer due to their influence on carcinogenesis and their possible association becomes a key element for future research. The main objective of this work was to identify Single Nucleotide Polymorphisms not described in the literature in the polymorphic regions of the NRF2 and KEAP1 genes in 40 patients with histologically confirmed hormone-dependent breast cancer and to establish a possible association with the histological grade of the tumour. To identify the polymorphisms under study, PCR and Sanger sequencing were used. In order to be able to relate and compare the results obtained in the blood with the tumour tissue, the technique of extracting gDNA from mouse paraffin-embedded tissue was optimized, using 9 samples of mouse mammary tissue. Polymorphisms rs6721961, rs6706649, rs35652124 from NRF2 and rs1048290 from KEAP1 were identified in all samples, and de novo mutations were not identified. These results emphasize the need to study the tumour tissue and verify if it presents alterations in gene expression and appearance of new tumour-related mutations. The gDNA extraction from paraffin-embedded tissue samples from mouse mammary sections was successful, so the protocol was properly optimized, allowing it to be replicated, in future, in human samples. |
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NRF2 and Keap1 Genetic Polymorphisms in Breast CancerCancro da MamaKeap1Nrf2Polimorfismos GenéticosSnpsDomínio/Área Científica::Ciências Médicas::Ciências BiomédicasCancer is one of the diseases with the highest mortality rate, with breast cancer being the most frequent and with the highest mortality rate in females. Human exposure to oxidative stress may be correlated with the development of several pathologies, including cancer. The pathways involved in oxidative stress represent a risk factor for the development of breast cancer. One of the most important factors in oxidative stress regulation is the NF-E2-related factor 2 (Nrf2)-Kelch-like ECH-associated protein 1 (Keap1) (Nrf2-Keap1) complex. The regulation of this complex may be compromised, due to somatic mutations that influence the loss of negative regulation or the presence of Single Nucleotide Polymorphisms in the promoter region of the NRF2 gene, which may be related to changes in the expression of this transcription factor and its biological consequences. Genetic polymorphisms have become potential research targets in breast cancer due to their influence on carcinogenesis and their possible association becomes a key element for future research. The main objective of this work was to identify Single Nucleotide Polymorphisms not described in the literature in the polymorphic regions of the NRF2 and KEAP1 genes in 40 patients with histologically confirmed hormone-dependent breast cancer and to establish a possible association with the histological grade of the tumour. To identify the polymorphisms under study, PCR and Sanger sequencing were used. In order to be able to relate and compare the results obtained in the blood with the tumour tissue, the technique of extracting gDNA from mouse paraffin-embedded tissue was optimized, using 9 samples of mouse mammary tissue. Polymorphisms rs6721961, rs6706649, rs35652124 from NRF2 and rs1048290 from KEAP1 were identified in all samples, and de novo mutations were not identified. These results emphasize the need to study the tumour tissue and verify if it presents alterations in gene expression and appearance of new tumour-related mutations. The gDNA extraction from paraffin-embedded tissue samples from mouse mammary sections was successful, so the protocol was properly optimized, allowing it to be replicated, in future, in human samples.O cancro é uma das doenças com maior taxa de mortalidade, sendo o cancro da mama o mais frequente e com maior taxa de mortalidade no sexo feminino. A exposição do ser humano ao stress oxidativo pode estar correlacionada com o desenvolvimento de diversas patologias, entre elas o cancro. As vias envolvidas no stress oxidativo representam um fator de risco para o desenvolvimento do cancro da mama. Um dos fatores mais importantes na regulação do stress oxidativo é o complexo NF-E2-related factor 2 (Nrf2)-Kelch-like ECH-associated protein 1 (Keap1) (Nrf2-Keap1). A regulação deste complexo pode estar comprometida devido a mutações somáticas que influenciem a perda da regulação negativa ou à presença de Single Nucleotide Polymorphisms na região promotora do gene NRF2, que podem estar relacionados com alterações na expressão deste fator de transcrição e respetivas consequências biológicas. Os polimorfismos genéticos tornaram-se potenciais alvos de investigação no cancro da mama devido à influência que os mesmos podem representar na carcinogénese e a sua possível associação torna-se um elemento-chave para investigação futura. O objetivo principal deste trabalho consistiu na identificação de Single Nucleotide Polymorphisms não descritos na literatura nas regiões polimórficas dos genes NRF2 e KEAP1 em 40 doentes com cancro da mama confirmado histologicamente como hormonodependente e estabelecer uma possível associação com o grau histológico do tumor. Para a identificação dos polimorfismos em estudo usou-se PCR e sequenciação de Sanger. De forma a poder relacionar e comparar os resultados obtidos no sangue com o tecido tumoral, foi otimizada a técnica de extração de gDNA de tecido parafinado de rato, usando para tal 9 amostras de tecido mamário de rato. Os polimorfismos rs6721961, rs6706649, rs35652124 do NRF2 e rs1048290 da KEAP1 foram identificados em todas as amostras, não tendo sido identificadas mutações de novo. Estes resultados enfatizam a necessidade de estudar o tecido tumoral e verificar se este apresenta alterações quer a nível da expressão de genes como a nível do aparecimento de novas mutações relacionadas com o tumor. A extração de gDNA de amostras de tecido parafinado de cortes mamários de rato foi bemsucedida pelo que o protocolo foi devidamente otimizado, permitindo que o mesmo seja replicado, no futuro, em amostras humanas.Granadeiro, Luiza Augusta Tereza Gil BreitenfeldPatricio, Ana Cristina Ramalhinho TavaresAlmeida, Micaela Carina PereirauBibliorumCosta, Diana Raquel Careano da2023-02-10T16:38:19Z2022-11-222022-10-102022-11-22T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10400.6/12896TID:203219945enginfo: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:RCAAP2023-12-15T09:56:16Zoai:ubibliorum.ubi.pt:10400.6/12896Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T00:52:23.966299Repositório 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 |
NRF2 and Keap1 Genetic Polymorphisms in Breast Cancer |
title |
NRF2 and Keap1 Genetic Polymorphisms in Breast Cancer |
spellingShingle |
NRF2 and Keap1 Genetic Polymorphisms in Breast Cancer Costa, Diana Raquel Careano da Cancro da Mama Keap1 Nrf2 Polimorfismos Genéticos Snps Domínio/Área Científica::Ciências Médicas::Ciências Biomédicas |
title_short |
NRF2 and Keap1 Genetic Polymorphisms in Breast Cancer |
title_full |
NRF2 and Keap1 Genetic Polymorphisms in Breast Cancer |
title_fullStr |
NRF2 and Keap1 Genetic Polymorphisms in Breast Cancer |
title_full_unstemmed |
NRF2 and Keap1 Genetic Polymorphisms in Breast Cancer |
title_sort |
NRF2 and Keap1 Genetic Polymorphisms in Breast Cancer |
author |
Costa, Diana Raquel Careano da |
author_facet |
Costa, Diana Raquel Careano da |
author_role |
author |
dc.contributor.none.fl_str_mv |
Granadeiro, Luiza Augusta Tereza Gil Breitenfeld Patricio, Ana Cristina Ramalhinho Tavares Almeida, Micaela Carina Pereira uBibliorum |
dc.contributor.author.fl_str_mv |
Costa, Diana Raquel Careano da |
dc.subject.por.fl_str_mv |
Cancro da Mama Keap1 Nrf2 Polimorfismos Genéticos Snps Domínio/Área Científica::Ciências Médicas::Ciências Biomédicas |
topic |
Cancro da Mama Keap1 Nrf2 Polimorfismos Genéticos Snps Domínio/Área Científica::Ciências Médicas::Ciências Biomédicas |
description |
Cancer is one of the diseases with the highest mortality rate, with breast cancer being the most frequent and with the highest mortality rate in females. Human exposure to oxidative stress may be correlated with the development of several pathologies, including cancer. The pathways involved in oxidative stress represent a risk factor for the development of breast cancer. One of the most important factors in oxidative stress regulation is the NF-E2-related factor 2 (Nrf2)-Kelch-like ECH-associated protein 1 (Keap1) (Nrf2-Keap1) complex. The regulation of this complex may be compromised, due to somatic mutations that influence the loss of negative regulation or the presence of Single Nucleotide Polymorphisms in the promoter region of the NRF2 gene, which may be related to changes in the expression of this transcription factor and its biological consequences. Genetic polymorphisms have become potential research targets in breast cancer due to their influence on carcinogenesis and their possible association becomes a key element for future research. The main objective of this work was to identify Single Nucleotide Polymorphisms not described in the literature in the polymorphic regions of the NRF2 and KEAP1 genes in 40 patients with histologically confirmed hormone-dependent breast cancer and to establish a possible association with the histological grade of the tumour. To identify the polymorphisms under study, PCR and Sanger sequencing were used. In order to be able to relate and compare the results obtained in the blood with the tumour tissue, the technique of extracting gDNA from mouse paraffin-embedded tissue was optimized, using 9 samples of mouse mammary tissue. Polymorphisms rs6721961, rs6706649, rs35652124 from NRF2 and rs1048290 from KEAP1 were identified in all samples, and de novo mutations were not identified. These results emphasize the need to study the tumour tissue and verify if it presents alterations in gene expression and appearance of new tumour-related mutations. The gDNA extraction from paraffin-embedded tissue samples from mouse mammary sections was successful, so the protocol was properly optimized, allowing it to be replicated, in future, in human samples. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-11-22 2022-10-10 2022-11-22T00:00:00Z 2023-02-10T16:38:19Z |
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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masterThesis |
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publishedVersion |
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http://hdl.handle.net/10400.6/12896 TID:203219945 |
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eng |
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