Evidence for estrogen receptor-mediated regulation of human gonadotropin-releasing hormone promoter activity in human placental cells

Two fragments of the human gonadotropin-releasing hormone (GnRH) promoter, one containing 0.6 kb of the downstream promoter sequence (H-1) and another 1.8 kb fragment (H-2) containing the upstream promoter region with a deletion of the downstream promoter sequence, were fused to a promoterless luciferase (Luc) reporter construct and transfected into the human placental (JEG) cells. JEG cells transfected with both constructs showed insignificant changes in luciferase activity in response to estradiol. However, cotransfection of H-2-Luc construct with a vector expressing a human estrogen receptor (ER) cDNA results in dose-dependent decreases in luciferase activity in response to estradiol. This ER mediated down-regulation of promoter activity was retained in constructs with the GnRH promoter deleted to position 548 bp 5' to the upstream transcription start site. Further deletion of upstream promoter sequence to 169 bp reversed the estrogen responsiveness from inhibitory to stimulatory. Thus, this study demonstrated that the upstream GnRH promoter region can be regulated by estrogen in transfected JEG cells.