Impact of early centrosome deregulation in malignant transformation

Detalhes bibliográficos
Autor(a) principal: Gomes, Inês Simões
Data de Publicação: 2021
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10451/53762
Resumo: Tese de mestrado, Biologia Molecular e Genética, Universidade de Lisboa, Faculdade de Ciências, 2022
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spelling Impact of early centrosome deregulation in malignant transformationCancro humanoEsófago de BarrettAmplificação do centrossomaInvasãoProgressão do tumorTeses de mestrado - 2022Departamento de Biologia VegetalTese de mestrado, Biologia Molecular e Genética, Universidade de Lisboa, Faculdade de Ciências, 2022Centrosome amplification (CA) is a hallmark of human cancers and is therefore a potential feature to explore for prognosis and therapy. However, the poor understanding of its origin and impact has limited its use in the clinic. Here, we used Barrett’s esophagus (BE) tumorigenesis as a human cancer model to test if CA has a contributory role in tumorigenesis and progression. Barrett’s esophagus is a premalignant condition and its neoplastic progression is a process with well-characterized and defined steps that go from metaplasia (premalignant condition - BE) to dysplasia (low- or high-grade intraepithelial neoplasia), adenocarcinoma (invasive neoplasia) and metastasis. Given that CA arises in BE metaplasia and that its incidence significantly increases from metaplasia to dysplasia, we posed the hypothesis that CA may play an important role in the acquisition of malignant properties. If this hypothesis is correct, then an increase of CA in metaplasia and/or a decrease of CA in dysplasia would be sufficient to respectively promote or reduce malignant properties such as invasiveness potential. To test these hypotheses, we first tested two different methods to reduce the CA levels in dysplasia cells: depletion of important molecules in the centrosome duplication cycle - PLK4, SAS6 and STIL - by siRNA, and inhibition of PLK4 activity using the specific chemical inhibitor centrinone-B. We then assessed changes in migration and invasive potential using 2D migration and invasion assays and 3D cell cultures. Both strategies were effective in reducing CA in dysplasia cells but as the treatment with the inhibitor acts directly on the protein's activity level, its effects were more quickly detected. Importantly, by providing the opportunity to reduce CA levels by affecting different molecules, the siRNA approach allowed us to confirm if the effects in the invasiveness properties of dysplasia cells is in fact a consequence of the reduction in the number of centrioles, and not just due to the dysfunction of specific molecules. Indeed, while results obtained with 2D migration and invasion assay were mostly inconclusive and further optimization is needed, we found that reduction of CA levels, with both approaches, reduced the invasiveness capacity of dysplasia cells in 3D cultures. Our findings therefore support a role for CA in promoting the invasiveness capacity in dysplasia, and provided important clues into their mechanisms that will be explored in the future. We then tested if an increase in CA is sufficient to induce invasion in metaplasia cells. To do this we first depleted p53 in metaplasia cells by siRNA, already known to increase CA in these cells, and then assessed their behavior using 3D cell cultures. Interestingly, our preliminary tests did not reveal any phenotypic differences in metaplasia cells with increased CA when compared to the controls. These findings indicate that increasing CA levels in metaplasia may not be sufficient, by itself, to induce an invasive capacity in metaplasia cells. In the future, testing different approaches to induce CA in these cells, as well as assessing their impact in cell migration and invasion using other assays, will be important to confirm this. By showing that CA contributes to the invasiveness potential in dysplasia, our results reveal the importance of CA in the acquisition of malignant properties in BE progression. These findings may contribute to new clinical tools, namely using CA as a biomarker of progression. Given widespread occurrence of CA in human tumors, our results may be extended to other cancers where CA is prevalent.Lopes, Carla A. M.Repositório da Universidade de LisboaGomes, Inês Simões202220212024-12-27T00:00:00Z2022-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10451/53762TID:202994546enginfo:eu-repo/semantics/embargoedAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-11-08T16:59:52Zoai:repositorio.ul.pt:10451/53762Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T22:04:43.400121Repositório 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 Impact of early centrosome deregulation in malignant transformation
title Impact of early centrosome deregulation in malignant transformation
spellingShingle Impact of early centrosome deregulation in malignant transformation
Gomes, Inês Simões
Cancro humano
Esófago de Barrett
Amplificação do centrossoma
Invasão
Progressão do tumor
Teses de mestrado - 2022
Departamento de Biologia Vegetal
title_short Impact of early centrosome deregulation in malignant transformation
title_full Impact of early centrosome deregulation in malignant transformation
title_fullStr Impact of early centrosome deregulation in malignant transformation
title_full_unstemmed Impact of early centrosome deregulation in malignant transformation
title_sort Impact of early centrosome deregulation in malignant transformation
author Gomes, Inês Simões
author_facet Gomes, Inês Simões
author_role author
dc.contributor.none.fl_str_mv Lopes, Carla A. M.
Repositório da Universidade de Lisboa
dc.contributor.author.fl_str_mv Gomes, Inês Simões
dc.subject.por.fl_str_mv Cancro humano
Esófago de Barrett
Amplificação do centrossoma
Invasão
Progressão do tumor
Teses de mestrado - 2022
Departamento de Biologia Vegetal
topic Cancro humano
Esófago de Barrett
Amplificação do centrossoma
Invasão
Progressão do tumor
Teses de mestrado - 2022
Departamento de Biologia Vegetal
description Tese de mestrado, Biologia Molecular e Genética, Universidade de Lisboa, Faculdade de Ciências, 2022
publishDate 2021
dc.date.none.fl_str_mv 2021
2022
2022-01-01T00:00:00Z
2024-12-27T00:00:00Z
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
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10451/53762
TID:202994546
url http://hdl.handle.net/10451/53762
identifier_str_mv TID:202994546
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/embargoedAccess
eu_rights_str_mv embargoedAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
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reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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