Detecção e expressão dos genes supressores p53 E c-Myc em tumores palpebrais de cães
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Data
2008-08-22
Autores
Lopes, Rodrigo Antonio [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
Os objetivos deste estudo foram detectar a presença e a expressão dos genes supressores p53 e c-Myc em tumores palpebrais, pelas técnicas de PCR, RTPCR, PCR-ELISA E RT-PCR-ELISA que até o então não foram descritas nestes tumores e nesta espécie. Foram utilizadas 10 amostras de tumores que foram fixados em formol e incluídos em parafina. O material foi obtido junto aos arquivos do Serviço de Patologia Veterinária, sendo nove amostras de tumores localizados nas pálpebras e terceira pálpebra e uma de tumor mamário para controle. Todos os tumores tiveram o seu diagnóstico firmado empregando-se a coloração de H.E e imunoistoquímica para citoqueratina AE1/AE3 e vimentina (V9), marcadores de tecido epitelial e mesenquimal, respectivamente. Os resultados indicaram que os tumores palpebrais e da terceira pálpebra aqui estudados verificou-se a presença do gene supressor p53 em 8 amostras (88,8%, n=8), e entre as amostras positivas (n=8), ele esteve expresso em 75 % delas. O gene supressor c-Myc esteve presente em 5 amostras (55,5%) e com expressão em 100% delas (n=5). Foi possível concluir que os tumores palpebrais e da terceira pálpebra de cães expressam o p-53 e c-Myc identificados pelas técnicas de PCR e RT-PCR, no entanto, as técnicas de PCR ELISA e RT-PCR ELISA foram mais importantes para avaliação da presença e expressão do oncogenes estudados, pois permitiram identificar produtos amplificados que não foram visualizados em gel de agarose.
The aims of this study were to detect the presence and the expression of p53 and c-Myc suppressor genes in eyelid tumors of dogs, by the PCR, RT-PCR, PCR-ELISA and RT-PCR-ELISA techniques, which until then they were not described in these tumors and in this specie. Ten samples of tumors were fixed in formalin and included in paraffin. The material was obtained from the archives of the Department of Veterinary Pathology, being nine samples of epithelial tumors located in the eyelids and the third eyelid, and a breast tumor which was used as a positive control of the reactions. All the samples had reached their diagnosis employing up the HE technique, and the immunohistochemistry for cytokeratin AE1/AE3 and vimentin (V9). The results showed that the eyelid and the third eyelid tumors, here studied, (88.8%, n=8) of them demonstrated the presence of the p53 gene and between the positive samples (n=8), the expression was around 75%. The c-Myc gene was present in 55.5% (n=5) of the samples, with 100% of expression (n=5). Thus, it was possible to conclude that the eyelid and the third eyelid tumors of dogs express the p53 and c-Myc genes, identified by the techniques of PCR and RT-PCR, however, the PCR ELISA and RT-PCR ELISA techniques were of extreme importance for assessing the presence and expression of these studied genes, and they allowed to identify amplified products that were not visible on the electrophoresis on the agarose gel.
The aims of this study were to detect the presence and the expression of p53 and c-Myc suppressor genes in eyelid tumors of dogs, by the PCR, RT-PCR, PCR-ELISA and RT-PCR-ELISA techniques, which until then they were not described in these tumors and in this specie. Ten samples of tumors were fixed in formalin and included in paraffin. The material was obtained from the archives of the Department of Veterinary Pathology, being nine samples of epithelial tumors located in the eyelids and the third eyelid, and a breast tumor which was used as a positive control of the reactions. All the samples had reached their diagnosis employing up the HE technique, and the immunohistochemistry for cytokeratin AE1/AE3 and vimentin (V9). The results showed that the eyelid and the third eyelid tumors, here studied, (88.8%, n=8) of them demonstrated the presence of the p53 gene and between the positive samples (n=8), the expression was around 75%. The c-Myc gene was present in 55.5% (n=5) of the samples, with 100% of expression (n=5). Thus, it was possible to conclude that the eyelid and the third eyelid tumors of dogs express the p53 and c-Myc genes, identified by the techniques of PCR and RT-PCR, however, the PCR ELISA and RT-PCR ELISA techniques were of extreme importance for assessing the presence and expression of these studied genes, and they allowed to identify amplified products that were not visible on the electrophoresis on the agarose gel.
Descrição
Palavras-chave
Pálpebras - Tumores, Teste imunoenzimático, Polimerase - Reação em cadeia, Eyelids - Neoplasms
Como citar
LOPES, Rodrigo Antonio. Detecção e expressão dos genes supressores p53 E c-Myc em tumores palpebrais de cães. 2008. 68 f. Dissertação (mestrado) - Universidade Estadual Paulista, Faculdade de Odontologia, 2008.