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Mémoire de Maîtrise
DOI
https://doi.org/10.11606/D.42.2007.tde-18102007-144836
Document
Auteur
Nom complet
Dayson Friaça Moreira
Unité de l'USP
Domain de Connaissance
Date de Soutenance
Editeur
São Paulo, 2007
Directeur
Jury
Belizario, Jose Ernesto
Malpartida, Humberto Miguel Garay
Seelaender, Marilia Cerqueira Leite
Titre en portugais
Redução do crescimento e resistência celular de carcinoma mamário após silenciamento do gene PIF/DCD (Proteolysis-Inducing Factor) via shRNAi.
Mots-clés en portugais
Caquexia
Dermicidina
Fator indutor de proteólise
RNA de interferência
Resumé en portugais
A expressão do PIF/DCD em tumores tem sido relacionada ao crescimento e resistência à morte celular e à indução de caquexia em camundongos. O objetivo deste estudo foi investigar o papel do PIF/DCD nestes processos. Foram construídos três PIF/DCD-RNAi chamados de IBC-I, II, III e pKLO (controle), compreendendo diferentes áreas do mRNA, geramos clones celulares PIF/DCD-RNAi e comparamos a expressão do mRNA por RT-PCR em tempo real. Depois, foi avaliado o crescimento e formação de colônias dos clones RNAi in vitro e a progressão tumoral in vivo em camundongo Nude. Os RNAi IBC-I, II e III reduziram a expressão em 93,25% dos transcritos do PIF/DCD, alem de reduzir em 61,7% a capacidade de formação de colônias em comparação ao controle (p<0.001). Nos experimentos in vivo, os camundongos 2 meses do grupo pKLO tiveram um retardo no desenvolvimento muscular. O grupo pKLO de 8 meses apresentou uma redução de 35% do seu peso. Ambos mostraram diminuição da massa muscular (p<0,05). Estes resultados confirmam o papel do PIF/DCD na progressão tumoral.
Titre en anglais
Reduction of growth and resistance cellular of mammary carcinoma after silencing of gene PIF/DCD (Proteolysis-inducing Factor) by shRNAi.
Mots-clés en anglais
Cachexia
Dermcidin
Proteolysis inducing-factor
RNA interference
Resumé en anglais
The PIF/DCD expression in tumors has been related to the growth and resistance to cellular death and induction of cachexia in mice. The aim of this study was to investigate the role of PIF/DCD in these processes. It was designed three PIF/DCD-RNAi named IBC-I, II and III and one control pKLO, comprising different areas of the mRNA. Next, it was generated PIF/DCD-RNAi cell clones and compared mRNA expression by RT-PCR real time. After that, we evaluated the growth and colony formation ability of RNAi clones in vitro and in vivo tumorogenicity in Nude mice. The IBC-I, II and III RNAi reduced in 93,25% the expression of transcripts for PIF/DCD. There was also a significant reduction (61,7%) in the colony formation compared to the control (p<0.001). In the in vivo experiments, the 2-month-old mice in the pKLO group had muscular development retardation. The 8-month-old pKLO group presented a 35% reduction on its weight. Both groups had shown muscular mass reduction (p<0,05). These results confirm the role of PIF/DCD in the tumoral progression.
 
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MM_TDE_2007_12.pdf (2.74 Mbytes)
Date de Publication
2007-10-19
 
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