Modulation of AP-1 mediated estrogenic response by ormeloxifene in rat uterus.
Ormeloxifene is a selective estrogen receptor modulator that exerts antiestrogenic effects and thereby inhibits growth in uterus. The present study was undertaken to examine the AP-1 protein interaction with AP-1 enhancer DNA elements in rat uterus in vivo and in vitro with a view to explore the modulation of estrogen action mediated via alternative pathway under the influence of ormeloxifene (Orm). In addition, the changes in expression of c-fos and c-jun transcription factors and mRNA expression of growth factor (IGF-1) were investigated with a view to assess the AP-1 mediated transcription. Ovariectomizedoung adult rats were administered with estradiol-17beta (5 microg/100 g body weight) or Orm (200 microg/100 g body weight) or vehicle for 3 days and sacrificed on fourth day. Electrophoretic mobility shift assay using uterine nuclear fraction from various treatment groups demonstrated that Orm caused a significant reduction in E2 induced AP-1 DNA binding. In vitro study revealed that Orm promotes AP-1 complex formation whereas its 7-hydroxy derivative inhibits it significantly. Uterine expression of c-fos and c-jun was increased significantly in Orm treated rats as compared to vehicle treated rats. However, the expression of c-fos and c-jun was decreased in rats receiving Orm plus E2. Semi-quantitative RT-PCR analysis revealed that mRNA expression of IGF-1 was increased in E2 treated group as compared to control group whereas reduced expression was observed in Orm treated rats as compared to E2 treated rats. The uterine weight and IGF-1 mRNA showed similar pattern, indicating that IGF-1 is involved in regulation of uterine weight. These results indicate that 7-hydroxy ormeloxifene (an active metabolite of Orm) is a potent antagonist at AP-1 sites. It inhibits the function of AP-1 transcription factors rather than their expression as evident by downregulation of mRNA expression of AP-1 regulated gene IGF-1, thereby inhibits proliferation in rat uterus. Study suggested a non-classicalregulation of estrogen action on uterus by ormeloxifene.
Awasthi
Smita
S
Division of Endocrinology, Central Drug Research Institute, Lucknow 226001, India.
Daverey
Amita
A
Dwivedi
Anila
A
D000818:Animals; D001578:Benzopyrans / Q000494:pharmacology*; D001665:Binding Sites; D004247:DNA / Q000378:metabolism; D020847:Estrogen Receptor Modulators / Q000494:pharmacology; D004967:Estrogens / Q000494:pharmacology*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D016762:Genes, fos; D016758:Genes, jun; D007334:Insulin-Like Growth Factor I / Q000235:genetics; D009929:Organ Size / Q000187:drug effects; D011485:Protein Binding; D051381:Rats; D017207:Rats, Sprague-Dawley; D018808:Transcription Factor AP-1 / Q000378:metabolism*; D014599:Uterus / Q000033:anatomy & histology / Q000187:drug effects* / Q000378:metabolism
D001578:Benzopyrans; D020847:Estrogen Receptor Modulators; D004967:Estrogens; D018808:Transcription Factor AP-1; C508548:ormeloxifene; D007334:Insulin-Like Growth Factor I; D004247:DNA