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Master's Dissertation
DOI
https://doi.org/10.11606/D.42.2018.tde-13032018-094101
Document
Author
Full name
Maria Aparecida de Oliveira
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2017
Supervisor
Committee
Santelli, Glaucia Maria Machado (President)
Amaral, Jonatas Bussador do
Barcelos, Denise
Debbio, Carolina Beltrame Del
Title in Portuguese
Aspectos moleculares e destino de células tumorais humanas submetidas a alterações de ploidia.
Keywords in Portuguese
Aneuploidia
Câncer
Citoesqueleto de actina
HIPPO
Instabilidade cromossômica
Abstract in Portuguese
A instabilidade cromossômica e a aneuploidia são características associadas às células malignas. Sabe-se que essas alterações podem ser resultantes de erros em eventos durante a mitose. Com o objetivo de gerar uma população com ganho de ploidia, utilizamos dois inibidores de fases distintas da mitose. Resultando no aumento da frequência celular em G2/M, ou seja, células com 4 vezes o número de cromossomos (4C) e de células com DNA acima de 4, hipertetraploide. Quantificamos a quantidade de núcleos e demonstramos que o tratamento, especificamente, levou a uma NCI, e não a multinucleação. Ambas linhagens celulares submetidas ao tratamento apresentaram alterações morfológicas, como protrusões de membrana, indicando alterações no citoesquelto. Nas análises de mRNA e da expressão proteica, observamos alterações na regulação da actina, coincidindo com a elevação dos mRNAs de YAP/TAZ, efetores co-transcricionais da via Hippo, regulada por alterações no citoesqueleto de actina. Desde modo, propomos, que o tratamento utilizado é um método eficiente para o estudo de células aneuploides e da NCI, que o citoesqueleto de actina é modulado por esse fenótipo e requer YAP/TAZ, provavelmente para manter a sobrevivência e favorecer a proliferação celular observada após o tratamento.
Title in English
Molecular aspects and fate of human tumor cells undergoing ploidy changes.
Keywords in English
Actin cytoskeleton
Aneuploidy
Cancer
Chromosomal instability
HIPPO
Abstract in English
Chromosomal instability and aneuploidy are characteristics associated with malignant cells. It is known that these changes may be due to errors in events during mitosis. In order to generate a population with gain of ploidy, we used two inhibitors of distinct phases of mitosis. Resulting in increasing cell frequency in G2/M, cells with 4 times the number of chromosomes (4C) and cells with DNA above 4, hypertetraploid. We quantified the number of nuclei and demonstrated that the treatment specifically led to NCI, not multinucleation. Both cell lines submitted to treatment presented morphological alterations, such as membrane protrusions, indicating changes in the cytoskeleton. In the analysis of mRNA and protein expression, we observed alterations in actin regulation, coinciding with the elevation of YAP / TAZ mRNAs, co-transcriptional effectors of the Hippo signaling pathway, regulated by changes in the actin cytoskeleton. We propose, that the treatment used is an efficient method for the study of aneuploid cells as well NCI. Also, the actin cytoskeleton is modulated by that phenotype which requires high YAP / TAZ, probably to maintain cell survival and promote cell proliferation observed.
 
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Publishing Date
2018-03-13
 
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