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Master's Dissertation
DOI
10.11606/D.42.2012.tde-26072012-102241
Document
Author
Full name
Felipe da Costa Souza
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2012
Supervisor
Committee
Strauss, Eugenia Costanzi (President)
Bajgelman, Marcio Chaim
Ventura, Armando Morais
Title in Portuguese
Geração e caracterização de linhagens isogênicas portadoras de mutantes de p53: modelo para avaliar a estratégia de reparação dos genes p53 e p16 INK4A na presença dos mutantes p53R175H e p53R248Q.
Keywords in Portuguese
Adenovírus
Ciclo celular
Cinética
Mutantes de P53
Neoplasias
Reparação de DNA
Abstract in Portuguese
A destruição funcional das vias de controle do ciclo celular constituem um evento comuns em todos os tumores humanos. Muitos estudos associam mutações em p53 com mau prognostico no tratamento do câncer. Nesse trabalho, visamos a geração e caracterização de linhagens isogênicas portando diferentes mutantes de p53 como modelo de estudo para remediação simultânea de p53 e p16 na presença de mutantes hotspots específicos. Os mutantes R175H e R248Q não geraram alterações na cinética de proliferação da linhagem H358, mas levaram a um aumento de 27,5% na eficiência de plaqueamento e, no caso de R248Q, ao dobro de eficiência na formação de colônias em suspensão. Os resultados do tratamento das linhagens isogênicas com adenovírus Adp16 e Adp53 mostraram que os mutantes não interferiram no parada do ciclo celular em G1 induzida por p16.
Title in English
Generation and characterization of isogenic cell lines harboring p53 mutants: a model for the evaluation of p53 and p16INK4A replacement in the presence of p53R175H and p53R248Q.
Keywords in English
Adenovirus
Cell cycle
DNA repair
Kinetics
Mutants of p53
Neoplasms
Abstract in English
Alterations of the cell cycle pathway are a common event in all human tumors. Several studies have shown a correlation between hotspot mutations and an unfavorable profile for cancer therapies. Hence, this study aims the generation and characterization of isogenic cell lines, harboring p53 mutants, as model to investigate the replacement of p53 and p16 genes on these mutant H358 cell lines. Our data identified that neither p53R175H nor p53R248Q mutants accelerated cell cycle progression. However, both leads to a 27,5% increased plate efficiency while R248Q leads to a two-fold increases in the number of colonies formed in soft agar. Our data also showed that the mutants did not affect the efficiency of p16 replacement.
 
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Publishing Date
2012-08-15
 
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