ABSTRACT
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.
Subject(s)
Chromosomes, Human, Pair 10/genetics , Exons/genetics , Genes, Homeobox/genetics , Homeodomain Proteins/genetics , Zebrafish Proteins , Animals , Base Sequence , Humans , In Situ Hybridization, Fluorescence , Introns/genetics , Mice , Molecular Sequence Data , RNA Splice Sites/genetics , Repressor Proteins , TATA Box/genetics , Transcription Factors , Untranslated Regions/geneticsSubject(s)
ATP-Binding Cassette Transporters/metabolism , Drug Resistance, Multiple , Hepatitis C, Chronic/metabolism , RNA, Messenger/analysis , ATP-Binding Cassette Transporters/genetics , DNA Primers/chemistry , Fluorescence , Fluorescent Antibody Technique, Direct , Hepatitis C, Chronic/genetics , Humans , Multidrug Resistance-Associated Proteins , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
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.
Subject(s)
Breast Neoplasms/metabolism , Neoplasm Proteins/metabolism , Ornithine Decarboxylase/metabolism , RNA, Messenger/metabolism , Female , Humans , In Situ Hybridization/methods , Proto-Oncogene Proteins c-myc/metabolism , Reverse Transcriptase Polymerase Chain Reaction/methods , Tumor Cells, CulturedABSTRACT
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.