EFFECT OF ESTRADIOL ON DHEAS PRODUCTION IN THE HUMAN ADRENOCORTICAL CELL-LINE, H295R

OBJECTIVE: To determine if estradiol regulates DHEA and DHEAS producti on in a human adrenocortical (H295R) cell line and to determine if thi s effect is receptor mediated. METHODS: NCI-H295 (H295R) cells were ri nsed and placed in phenol red free Dulbecco's Modified Eagle's-F-12 me dium supplemented with 0.1% charcoal-stripped serum. After 24 hours, c ells were rinsed and treated based on experimental design. The effects of estradiol were investigated by: 1) treatment of cells with increas ing concentrations of estradiol (300-3000 nmol/L) with or without fors kolin (10 mu mol/L), 2) treatment of cells with the nonsteroidal synth etic estrogen diethylstilbestrol (DES) (300-3000 nmol/L) with or witho ut forskolin (10 mu mol/L), and 3) treatment of cells with an estradio l antagonist (ICI 182,780) in the presence of estradiol. RESULTS: Estr adiol alone increased the based production of DHEAS in H295R cells in a concentration-dependent manner with a maximal effect at 1000 nmol/L. Forskolin treatment increased the basal production of DHEAS ten-fold. Estradiol also increased the forskolin stimulation of DHEAS productio n two-fold. In contrast, DES alone or DES in addition to forskolin did not stimulate DHEAS production. Estradiol, in contrast, inhibited H29 5R adrenal cell production of cortisol whereas DES exhibited a similar inhibition. The estrogen receptor antagonist ICI 182,780 was unable t o inhibit the stimulatory effect of estradiol. Finally, estradiol in a concentration-dependent manner suppressed 3 beta-hydroxysteroid dehyd rogenase (3 beta HSD) activity in H295R adrenal cells. CONCLUSION: The se experiments support the role of estradiol in regulating DHEAS produ ction by inhibiting 3 beta HSD activity; however, the mechanism appear s to require high concentrations of estradiol and appears to be indepe ndent of the estrogen receptor. Copyright (C) 1998 by the Society for Gynecologic Investigation.