RESUMO
We have studied the expression of ornithine decarboxylase (ODC) mRNA by in situ hybridization and in situ RT-PCR in human breast cancer MCF-7 cell line. In situ RT-PCR demonstrated the overexpression of ODC mRNA, localized over the cytoplasm, while a very low signal was detected by in situ hybridization. Our findings indicate that in situ RT-PCR could represent a useful tool to study different levels of ODC expression in normal and tumor tissues. Since ODC expression is regulated by c-Myc oncoprotein, this model could be useful to monitor in vivo the effects of new anti-neoplastic molecules, specific inhibitors of c-Myc.
Assuntos
Neoplasias da Mama/metabolismo , Proteínas de Neoplasias/metabolismo , Ornitina Descarboxilase/metabolismo , RNA Mensageiro/metabolismo , Feminino , Humanos , Hibridização In Situ/métodos , Proteínas Proto-Oncogênicas c-myc/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Células Tumorais CultivadasRESUMO
Expression of ornithine decarboxylase (ODC) is induced by c-Myc oncoprotein and is required for cell proliferation and tumour growth. We have studied the expression of ODC mRNA by in situ hybridisation and in situ RT-PCR in archival human hyperplastic breast tissues. A very low signal was detected by in situ hybridisation, while the in situ RT-PCR on human breast archival tissues demonstrated an over-expression of ODC mRNA in epithelial cells characterised by some degree of hyperplasia, maintaining the morphology of the archival tissue intact despite the multiple steps of fixation, permeabilization and thermal cycling.
Assuntos
Mama/enzimologia , Mama/patologia , Ornitina Descarboxilase/metabolismo , Células Epiteliais/enzimologia , Células Epiteliais/patologia , Humanos , Hiperplasia/enzimologia , Hiperplasia/genética , Hiperplasia/patologia , Hibridização In Situ , Ornitina Descarboxilase/genética , Inclusão em Parafina , Permeabilidade , RNA Mensageiro/biossíntese , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sensibilidade e Especificidade , Fixação de TecidosAssuntos
Transportadores de Cassetes de Ligação de ATP/metabolismo , Resistência a Múltiplos Medicamentos , Hepatite C Crônica/metabolismo , RNA Mensageiro/análise , Transportadores de Cassetes de Ligação de ATP/genética , Primers do DNA/química , Fluorescência , Técnica Direta de Fluorescência para Anticorpo , Hepatite C Crônica/genética , Humanos , Proteínas Associadas à Resistência a Múltiplos Medicamentos , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
In the present study, we report the genomic reconstruction of the human homeobox-containing gene HHEX by the use of the data available in public databases. This analysis allowed characterization of the gene organization showing that it is very similar to the mouse gene. Moreover the gene was mapped using FISH to 10q24.