Identificação e validação de genes diferencialmente expressos em carcinoma de pênis
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Data
2013-03-06
Autores
Mota, Mânlio Tasso de Oliveira [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
O carcinoma de pênis (CEP) é um tumor epitelial invasivo raro com alta morbidade decorrente da própria doença e/ou de seu tratamento. O perfil socioeconômico e cultural dos pacientes dificulta o diagnóstico precoce, tratament o e seguimento dos enfermos. Pacientes sem tratamento geralmente morrem dentro de dois anos após diagnóstico devido à proliferação celular regional ou a metástases distantes. Não há padrão nos sistemas de estadiamento e nas condutas clínicas, resultando em dificuldades na abordagem terapêutica. Comparado ao câncer cervical, poucos estudos moleculares em CEP foram realizados. O presente projeto teve como objetivo geral identificar genes diferencialmente expressos em tecidos penianos tumorais e normais e o possível papel do vírus do papiloma humano (HPV) no desenvolvimento de CEP. Diferenças na expressão gênica entre tecidos tumorais e normais foram verificadas pela metodologia de RaSH (rapid subctration hybridization), que selecionou 5 genes possivelmente rel acionados à carcinogênese ( KIAA1033, NAMPT, RPL6, CDKN2A e ANXA1) com expressão alterada em tecidos tumorais. A validação das alterações de expressão destes genes foi realizada por polymerase chain reaction (reação em cadeia da polimerase, PCR) em tempo real. Tanto os genes como as proteínas de ANXA1 como CDKN2A apresentaram aumento de expressão em pacientes com presença de HPV de alto risco em comparação com tecidos negativos para HPV. Por imunohistoquímica foi possível estabelecer correlação entre alterações na expressão da proteína anexina A1 e a presença de HPV de alto risco. Pacientes portadores de CEP provenientes de duas regiões (São Paulo e Pará) foram avaliados para a presença de HPV por PCR convencional. Amostras positivas foram genotipadas por hib ridização...
Penile carcinoma is a rare epithelial tumor with high morbidity due to own disease and/or treatment. The socio -economic and cultural profile of the patients hampers early diagnosis, treatment and follow-up. Without treat ment patients usually dies within two years after first diagnosis due to regional cellular proliferation or distant metastasis. There is no standard in staging systems or in clinical procedures, resulting in difficulties in the therapeutic approach. Compar ed to cervical cancer, there are few molecular studies about this tumor. The present work aimed to identify differentially expressed genes in tumor and normal penile tissues and the role of human papillomavirus (HPV) in the development of this tumor. Differences in gene expression between tumoral and normal tissues were accessed by RaSH (rapid subctration hybridization) methodology, which selected five possibly carcinogenesis related genes (KIAA1033, NAMPT, RPL6, CDKN2A and ANXA1) with altered expression in tumor tissues. The validation of gene expression changes was performed by real time polymerase chain reaction (PCR). The ANXA1 and CDKN2A, both genes and proteins, showed superexpression in patients with high-risk HPV compared to HPV negative tissues. Immunohistochemical assays established a correlation between alterations in the superexpression of annexin A1 and the presence of high risk HPV. Penile cancer harboring patients from two regions (São Paulo and Pará) was assessed for HPV presence by conventional PCR. Positive samples were genotyped by reverse hybridization-based line probe assay (INNO -LiPA). The overall HPV prevalence in single or multiple infections was significantly greater in Pará (81.67%) than in São Paulo (64.10%). A wilder range of genotypes were found in Pará, suggesting that the genotypes circulating in the population of Pará... (Complete abstract click electronic access below)
Penile carcinoma is a rare epithelial tumor with high morbidity due to own disease and/or treatment. The socio -economic and cultural profile of the patients hampers early diagnosis, treatment and follow-up. Without treat ment patients usually dies within two years after first diagnosis due to regional cellular proliferation or distant metastasis. There is no standard in staging systems or in clinical procedures, resulting in difficulties in the therapeutic approach. Compar ed to cervical cancer, there are few molecular studies about this tumor. The present work aimed to identify differentially expressed genes in tumor and normal penile tissues and the role of human papillomavirus (HPV) in the development of this tumor. Differences in gene expression between tumoral and normal tissues were accessed by RaSH (rapid subctration hybridization) methodology, which selected five possibly carcinogenesis related genes (KIAA1033, NAMPT, RPL6, CDKN2A and ANXA1) with altered expression in tumor tissues. The validation of gene expression changes was performed by real time polymerase chain reaction (PCR). The ANXA1 and CDKN2A, both genes and proteins, showed superexpression in patients with high-risk HPV compared to HPV negative tissues. Immunohistochemical assays established a correlation between alterations in the superexpression of annexin A1 and the presence of high risk HPV. Penile cancer harboring patients from two regions (São Paulo and Pará) was assessed for HPV presence by conventional PCR. Positive samples were genotyped by reverse hybridization-based line probe assay (INNO -LiPA). The overall HPV prevalence in single or multiple infections was significantly greater in Pará (81.67%) than in São Paulo (64.10%). A wilder range of genotypes were found in Pará, suggesting that the genotypes circulating in the population of Pará... (Complete abstract click electronic access below)
Descrição
Palavras-chave
Câncer, Penis - Câncer, Penis - Tumores, Carcinoma de celulas escamosas, Virus do papiloma, Marcadores biologicos de tumor
Como citar
MOTA, Mânlio Tasso de Oliveira. Identificação e validação de genes diferencialmente expressos em carcinoma de pênis. 2013. 101 f. Tese (doutorado) - Universidade Estadual Paulista Julio de Mesquita Filho, Instituto de Biociências, Letras e Ciências Exatas de São José do Rio Preto, 2013.