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Previous issue date: 2015-02-04 / The Cortical dysplasia is one of the most frequent forms of malformations of cortical development, with a condition that causes a large portion of refractory partial epilepsy to drug treatment. This condition is characterized by a heterogeneous group of cortical lesions and can also be described as cortical dysgenesis or neuronal migration disorder. The genetic factors involved in cortical dysplasia, such as the expression of the telomerase enzyme, are not commonly investigated, mainly due to the limited number of cases is the lack of an experimental model. Objective: This study aimed to analyze the activity and the expression of the enzyme telomerase in patients with cortical dysplasia of Taylor, normal fibroblasts, stem cells from umbilical cord tissue and human tumor. Methodology: the activity and expression of hTERT portion of telomerase were evaluated in patient fibroblasts with Cortical Dysplasia of Taylor (n = 1) and compared with control patients fibroblasts (n = 3) and umbilical cord stem cells (n = 3) and human tumor cell line A549 were maintained in culture. The analysis of the hTERT gene was done by PCR and the telomerase activity was measured by TRAP assay that uses real-time PCR. Results: telomerase activity was higher in tumor cells (Ct = 28.32), while in other cell types and other types studied values were similar: control (Ct = 33.73) Umbilical Cord (Ct = 32.88) and dysplasia (Ct = 33.56). The expression of hTERT portion of telomerase was the control group was 2,58x more expressed, the Dysplasia and tumor groups were 61,25x and 635,48x more expressed respectively as the control. Conclusion: The telomerase appeared to be more active and higher expression in the tumor cell line when compared to other cell types surveyed. The hTERT gene was more expressed in the patient Taylor cortical dysplasia than in the other groups and control umbilical cord indicating that telomerase is expressed in most patients with this disease. / A Displasia Cortical ? uma das formas mais frequentes de malforma??es do desenvolvimento cortical, sendo uma patologia que causa uma grande parcela de epilepsias parciais refrat?rias ao tratamento medicamentoso. Essa patologia ? caracterizada por um grupo heterog?nio de les?es corticais, podendo tamb?m ser descrita como disgenesia cortical ou desordem de migra??o neuronal. Os aspectos gen?ticos envolvidos na displasia cortical, como a express?o da enzima telomerase, n?o s?o comumente investigados, principalmente devido ao n?mero limitados de casos e a pela falta de um modelo experimental. Objetivo: este estudo teve como objetivo analisar a atividade e a express?o da enzima telomerase em pacientes com displasia cortical de Taylor, fibroblastos normais, c?lulas-tronco de tecido de cord?o umbilical e linhagem tumoral humana. Metodologia: a atividade e express?o da por??o hTERT da telomerase foram avaliadas em fibroblastos de paciente com Displasia Cortical de Taylor (n=1) e comparada com fibroblastos de pacientes do grupo controle (n=3) e c?lulas-tronco de cord?o umbilical (n=3) e a linhagem tumoral A549 humanos que foram mantidas em cultura. A an?lise do gene hTERT foi feita por PCR e a atividade da telomerase foi medida pelo ensaio TRAP que utiliza a PCR em tempo real. Resultados: a atividade da telomerase foi maior nas c?lulas tumorais (Ct=28,32), ao passo que nos demais tipos celulares e nos demais tipos estudados os valores foram semelhantes: Controle (Ct=33,73) Cord?o Umbilical (Ct=32,88) e Displasia (Ct=33,56). A express?o da por??o hTERT da telomerase foi do grupo Controle foi 2,58x mais expresso, os grupos Dsiplasia e Tumor estavam 61,25x e 635,48x mais expressos, respectivamente que o Controle. Conclus?o: a telomerase se mostrou mais ativa e com maior express?o na linhagem tumoral do que quando comparada com os outros tipos celulares pesquisados. O gene hTERT e mostrou mais expresso, no paciente com Displasia Cortical de Taylor do que nos outros grupos Controle e Cord?o Umbilical, indicando que a telomerase ? mais expressa em pacientes com esta patologia.
| Identifer | oai:union.ndltd.org:IBICT/oai:tede2.pucrs.br:tede/6214 |
| Date | 04 February 2015 |
| Creators | Borges, Juliano Viana |
| Contributors | Machado, Denise Cantarelli |
| Publisher | Pontif?cia Universidade Cat?lica do Rio Grande do Sul, Programa de P?s-Gradua??o em Gerontologia Biom?dica, PUCRS, Brasil, Instituto de Geriatria e Gerontologia |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da PUC_RS, instname:Pontifícia Universidade Católica do Rio Grande do Sul, instacron:PUC_RS |
| Rights | info:eu-repo/semantics/openAccess |
| Relation | 4438661476953179033, 600, 600, 600, 2296420844541114010, -969369452308786627 |
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