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-17 beta (5 mu g/100 g body weight) or Orm (200 mu g/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 E-2 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 E-2. Semi-quantitative RT-PCR analysis revealed that mRNA expression of IGF-1 was increased in E-2 treated group as compared to control group whereas reduced expression was observed in Orm treated rats as compared to E-2 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. (c) 2007 Published by Elsevier Ltd.