Análise da expressão e do número de cópias gênicas de microrna da região 8q24.21 no câncer gástrico
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Data
2018-05-24
Autores
Pereira, Ana Carolina Anauate [UNIFESP]
Orientadores
Smith, Marilia de Arruda Cardoso [UNIFESP]
Tipo
Dissertação de mestrado
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Objetivo: Analisar a expressão de miR1204, miR1205, miR1206, miR1207 e miR1208 em tecido gástrico neoplásico e não neoplásico de pacientes com e sem CG, determinar os genes de referência para essa análise, avaliar o número de cópias gênicas desses miRNA, e associar estas variáveis com os parâmetros clínicos e patológicos dos pacientes com CG. Métodos: A expressão dos miRNA alvos miR1204, miR1205, miR1206, miR12075p, miR12073p, e miR1208 foi avaliada por RTqPCR em 65 pares de amostras neoplásicas (T) e margem adjacente não neoplásicas (A) de pacientes com CG, e 20 amostras de tecido gástrico não neoplásica de indivíduos sem câncer submetidos à endoscopia (C), e determinada pelo método ΔCt. O número de cópias dos genes miR1204, miR1205, miR1207, e miR1208 foi determinado por qPCR em 69 amostras T e com o uso do programa Copy Caller. Resultados: O grupo de amostras C não diferiu em relação ao gênero e idade dos pacientes com CG (p>0,05). Apenas o miR1205 apresentou diferença de expressão entre os grupos de amostra A e C, com maior expressão no grupo A (p=0,02). Não foi observada expressão de miR1204 e miR1206 nas amostras de tecido gástrico analisadas. Foi observada uma correlação positiva e forte entre a expressão de miR12075p e miR1208 (p<0,01, ρ=0,71), e uma correlação positiva, mas fraca, entre a expressão de miR1205 e miR12073p (p=0,04, ρ=0,21). Não foi observada associação entre a expressão de miR1205, miR12075p, miR12073p, e miR1208 e o número de cópias dos seus respectivos genes (p>0,05). O aumento do número de cópias do gene miR1207 foi associado aos tumores localizados na região não cárdia do estômago (p=0,01), e a perda do número de cópias do gene miR1208 à presença de infecção por H. pylori (p=0,01). Conclusões: Não foram encontradas diferenças significantes entre a expressão de miR1205, miR12075p, miR12073p, e miR1208 nas amostras T vs A, e entre T vs C. Porém, o miR1205 apresentou expressão aumentada nas amostras A em relação às C. Os resultados sugerem que o miR1205 parece ter um papel importante no início do desenvolvimento do CG. Além disso, a ausência de associação entre a expressão desses miRNA e o número de cópias gênicas indica que a alteração do número de cópias do miR1205 não seja o mecanismo que leva à sua desregulação nas amostras avaliadas.
Objective: To analyze the expression of miR1204, miR1205, miR1206, miR1207 and miR1208 in neoplastic and nonneoplastic gastric tissue from patients with and without GC, determine the reference genes for this analysis, the number of gene copies of these miRNAs, and to associate these variables with the clinical and pathological parameters of GC patients. Methods: The expression of miR1204, miR1205, miR1206, miR12075p, miR12073p, and miR1208 target miRNA was evaluated by RTqPCR in 65 paired neoplastic (T) and adjacent nonneoplastic (A) samples from patients with GC, and 20 nonneoplastic gastric tissue samples from cancerfree individuals submitted to endoscopy (C), and determined by the ΔCt method. The number of copies of the miR1204, miR1205, miR1207, and miR1208 genes was determined by qPCR in 69 T samples using the Copy Caller program. Results: The C group of samples did not differ in relation to gender and age of CG patients (p> 0.05). Only miR1205 presented different expression between A and C groups, with higher expression in A group (p = 0.02). Expression of miR1204 and miR1206 was not observed in the analyzed gastric tissue samples. A strong positive correlation was observed between miR12075p and miR1208 expression (p <0.01, ρ = 0.71), and a very weak positive correlation between miR1205 and miR12073p expression (p = 0.04, ρ = 0.21). No association was observed between miR1205, miR12075p, miR12073p, and miR1208 expression and the number of copies of their respective genes (p> 0.05). Increased miR1207 gene copy number was associated with tumors located in noncardiac region of the stomach (p = 0.01), and the loss of copies of miR1208 gene was associated with H. pylori infection (p = 0.01). Conclusion: No significant differences were found between miR1205, miR12075p, miR12073p, and miR1208 expression in T vs A and T vs C samples. However, miR1205 showed an increased expression in A samples compared to C samples. The results suggest that miR1205 participates in the beginning of GC development. In addition, the absence of association between these miRNA expressions and the gene copy number indicates that the change in miR1205 copy number is not the mechanism leading to its deregulation in the samples tested.
Objective: To analyze the expression of miR1204, miR1205, miR1206, miR1207 and miR1208 in neoplastic and nonneoplastic gastric tissue from patients with and without GC, determine the reference genes for this analysis, the number of gene copies of these miRNAs, and to associate these variables with the clinical and pathological parameters of GC patients. Methods: The expression of miR1204, miR1205, miR1206, miR12075p, miR12073p, and miR1208 target miRNA was evaluated by RTqPCR in 65 paired neoplastic (T) and adjacent nonneoplastic (A) samples from patients with GC, and 20 nonneoplastic gastric tissue samples from cancerfree individuals submitted to endoscopy (C), and determined by the ΔCt method. The number of copies of the miR1204, miR1205, miR1207, and miR1208 genes was determined by qPCR in 69 T samples using the Copy Caller program. Results: The C group of samples did not differ in relation to gender and age of CG patients (p> 0.05). Only miR1205 presented different expression between A and C groups, with higher expression in A group (p = 0.02). Expression of miR1204 and miR1206 was not observed in the analyzed gastric tissue samples. A strong positive correlation was observed between miR12075p and miR1208 expression (p <0.01, ρ = 0.71), and a very weak positive correlation between miR1205 and miR12073p expression (p = 0.04, ρ = 0.21). No association was observed between miR1205, miR12075p, miR12073p, and miR1208 expression and the number of copies of their respective genes (p> 0.05). Increased miR1207 gene copy number was associated with tumors located in noncardiac region of the stomach (p = 0.01), and the loss of copies of miR1208 gene was associated with H. pylori infection (p = 0.01). Conclusion: No significant differences were found between miR1205, miR12075p, miR12073p, and miR1208 expression in T vs A and T vs C samples. However, miR1205 showed an increased expression in A samples compared to C samples. The results suggest that miR1205 participates in the beginning of GC development. In addition, the absence of association between these miRNA expressions and the gene copy number indicates that the change in miR1205 copy number is not the mechanism leading to its deregulation in the samples tested.
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Citação
PEREIRA, Ana Carolina Anauate. Análise da expressão e do número de cópias gênicas de microRNA da região 8q24.21 no câncer gástrico. São Paulo, 2018. 64 f. Dissertação (Mestrado em Biologia estrutural e funcional) - Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, 2018.