Effect of Estradiol on DHEAS Production in the Human Adrenocortical Cell Line,H295R

Abstract

Objective: To determine if estradiol regulates DHEA and DHEAS production in a human adrenocortical (H295R) cell line and to determine if this effect is receptor mediated.

Methods: NCI-H295 (H295R) cells were rinsed and placed in phenol red free Dulbecco's Modified Eagle's-F12 medium supplemented with 0.1% charcoal-stripped serum. After 24 hours, cells were rinsed and treated based on experimental design. The effects of estradiol were investigated by: 1) treatment of cells with increasing concentrations of estradiol (300-3000 nmol/L) with or without forskolin (10 μmol/L), 2) treatment of cells with the nonsteroidal synthetic estrogen diethylstilbestrol (DES) (300-3000 nmol/L) with or without forskolin (10 μmol/L), and 3) treatment of cells with an estradiol antagonist (ICI 182,780) in the presence of estradiol.

Results: stradiol alone increased the basal production 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 production two-fold. In contrast, DES alone or DES in addition to forskolin did not stimulate DHEAS production. Estradiol, in contrast, inhibited H295R adrenal cell production of cortisol whereas DES exhibited a similar inhibition. The estrogen receptor antagonist ICI 182,780 was unable to inhibit the stimulatory effect of estradiol. Finally, estradiol in a concentration-dependent manner suppressed 3β-hydroxysteroid dehydrogenase (3βHSD) activity in H295R adrenal cells.

Conclusion: These experiments support the role of estradiol in regulating DHEAS production by inhibiting 3βHSD activity; however, the mechanism appears to require high concentrations of estradiol and appears to be independent of the estrogen receptor.