Semiquantitative reverse transcription-polymerase chain reaction to evaluate the expression patterns of genes involved in the oestrogen pathway

被引:31
|
作者
Rey, JM
Pujol, P
Callier, P
Cavailles, V
Freiss, G
Maudelonde, T
Brouillet, JP
机构
[1] CHU Arnaud de Villeneuve, Lab Biol Cellulaire & Hormonale, F-34295 Montpellier 5, France
[2] Univ Montpellier 1, INSERM, U540, Unite Endocrinol Mol & Cellulaire Canc, F-34090 Montpellier, France
关键词
D O I
10.1677/jme.0.0240433
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
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 (ER alpha, ER beta), 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 ER alpha-positive and three ER alpha-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.
引用
收藏
页码:433 / 440
页数:8
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