Stimulation of endometrial glandular cells with genistein and daidzein and their effects on ERalpha- and ERbeta-mRNA and protein expresion.
Phytoestrogens seem to have estrogen-like effects in the human body as their structure is very similar to those estrogens produced in human glands. The aim of the present study was to analyse the effects of genistein and daidzein on estrogen receptor (ER)alpha- and ERbeta-mRNA and protein expression in the endometrium of premenopausal women.


METHODS
Glandular endometrial cells were isolated from endometrial biopsies obtained from regularly menstruating women undergoing gynaecological abrasio or hysterectomy. Cells were stimulated with single doses of genistein or daidzein. ERalpha- and ERbeta-protein expression were determined by immunocytochemical analysis. In addition ERalpha- and ERbeta-mRNA expression were determined by quantitative real-time RT-PCR. Quantification was carried out by the deltadelta C(T)-method using glyceraldehyde phosphate dehydrogenase (GAPDH) as housekeeping gene.


RESULTS
Endometrial glandular cells responded to stimulation with genistein and daidzein by alteration of both ERalpha- and ERbeta-mRNA expression. The effects were time- and dose-dependent. Detection of ERalpha- and ERbeta-protein expression by immunocytochemistry showed a dose-dependent regulation after stimulation of isolated endometrial cells with phytoestrogens.


CONCLUSIONS
According to our results, we suggest that ER expression in endometrial glandular cells is regulated by genistein and daidzein on the mRNA and protein levels. We could detect a decrease in ERalpha- and an increase in ERbeta-mRNA expression after stimulation with tested phytoestrogens. Our results are in line with findings that phytoestrogens act as anti-estrogens in organs expressing more ERalpha and as estrogens in ERbeta-presenting organs.
Staar
Stefanie
S
University of Rostock, Department of Obstetrics and Gynaecology, Faculty of Medicine, Doberaner Strasse 142, D-18055 Rostock, Germany.
Richter
Dagmar-Ulrike
DU
Makovitzky
Josef
J
Briese
Volker
V
Bergemann
Claudia
C
D001483:Base Sequence; D002478:Cells, Cultured; D017931:DNA Primers; D004717:Endometrium / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D047628:Estrogen Receptor alpha / Q000235:genetics / Q000378:metabolism*; D047629:Estrogen Receptor beta / Q000235:genetics / Q000378:metabolism*; D005260:Female; D019833:Genistein / Q000494:pharmacology*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D007529:Isoflavones / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017931:DNA Primers; D047628:Estrogen Receptor alpha; D047629:Estrogen Receptor beta; D007529:Isoflavones; D012333:RNA, Messenger; C004742:daidzein; D019833:Genistein; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases