title: Semiquantitative reverse transcription-polymerase chain reaction to evaluate the expression patterns of genes involved in the oestrogen pathway.
abstract: The increasing number of factors to be taken into account in the oestrogen transcriptional process has created a need to develop a rapid screening method to evaluate their role in physiology and pathology. Molecular biology techniques enable gene expression studies at the mRNA level with small amounts of tissues. We therefore developed a semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) technique using fluorescent oligonucleotides to analyse simultaneously a large panel of interrelated genes involved in the oestrogen transcriptional pathway using a moderately expressed housekeeping gene, the hypoxanthine phosphoribosyltransferase gene (HPRT), as the reference gene. Expression levels of oestrogen receptors (ERalpha, ERbeta), cofactors AIB1, RIP140, SMRT and the Fas-associated protein-tyrosine phophatase-1 (FAP-1) genes were evaluated in breast, endometrial and ovarian cancer cell lines and in three ERalpha-positive and three ERalpha-negative breast cancer tumours. This technique provides a rapid and reliable way to quantitate simultaneously numerous mRNAs of genes involved in the oestrogen pathway from small amounts of tissues.
authors.affiliation: Laboratoire de Biologie Cellulaire et Hormonale, CHU Arnaud de Villeneuve, 371 av. du Doyen G Giraud, 34295 Montpellier Cedex 5, France.
authors.forename: J M
authors.initials: JM
authors.lastname: Rey
authors.forename: P
authors.initials: P
authors.lastname: Pujol
authors.forename: P
authors.initials: P
authors.lastname: Callier
authors.forename: V
authors.initials: V
authors.lastname: Cavailles
authors.forename: G
authors.initials: G
authors.lastname: Freiss
authors.forename: T
authors.initials: T
authors.lastname: Maudelonde
authors.forename: J P
authors.initials: JP
authors.lastname: Brouillet
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000235:genetics; D004967:Estrogens / Q000378:metabolism*; D015870:Gene Expression; D051548:Histone Acetyltransferases; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D009687:Nuclear Proteins / Q000235:genetics; D056985:Nuclear Receptor Co-Repressor 2; D056920:Nuclear Receptor Coactivator 1; D000078723:Nuclear Receptor Interacting Protein 1; D054595:Protein Tyrosine Phosphatase, Non-Receptor Type 13; D017027:Protein Tyrosine Phosphatases / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D011960:Receptors, Estrogen / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014157:Transcription Factors / Q000235:genetics; D014407:Tumor Cells, Cultured
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; D017931:DNA Primers; D004268:DNA-Binding Proteins; D004967:Estrogens; C539130:NCOR2 protein, human; C000627127:NRIP1 protein, human; D009687:Nuclear Proteins; D056985:Nuclear Receptor Co-Repressor 2; D000078723:Nuclear Receptor Interacting Protein 1; D012333:RNA, Messenger; D011960:Receptors, Estrogen; D012097:Repressor Proteins; D014157:Transcription Factors; D051548:Histone Acetyltransferases; C538859:NCOA1 protein, human; D056920:Nuclear Receptor Coactivator 1; D007041:Hypoxanthine Phosphoribosyltransferase; C516742:PTPN13 protein, human; D054595:Protein Tyrosine Phosphatase, Non-Receptor Type 13; D017027:Protein Tyrosine Phosphatases