Effect of estradiol on DHEAS production in the human adrenocortical cell line, H295R.
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 mumol/L), 2) treatment of cells with the nonsteroidal synthetic estrogen diethylstilbestrol (DES) (300-3000 nmol/L) with or without forskolin (10 mumol/L), and 3) treatment of cells with an estradiol antagonist (ICI 182, 780) in the presence of estradiol.


RESULTS
Estradiol alone increased the basal 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 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 3 beta-hydroxysteroid dehydrogenase (3 beta HSD) activity in H295R adrenal cells.


CONCLUSIONS
These experiments support the role of estradiol in regulation DHEAS production by inhibiting 3 beta HSD activity; however, the mechanism appears to require high concentrations of estradiol and appears to be independent of the estrogen receptor.
Gell
J S
JS
Department of Obstetrics and Gynecology, University of Texas Southwestern Medical Center at Dallas 75235-9032, USA.
Oh
J
J
Rainey
W E
WE
Carr
B R
BR
D015096:3-Hydroxysteroid Dehydrogenases / Q000378:metabolism*; D000302:Adrenal Cortex / Q000187:drug effects / Q000378:metabolism*; D002460:Cell Line; D005576:Colforsin / Q000494:pharmacology; D003687:Dehydroepiandrosterone / Q000096:biosynthesis / Q000378:metabolism*; D019314:Dehydroepiandrosterone Sulfate / Q000378:metabolism*; D004054:Diethylstilbestrol / Q000494:pharmacology*; D004958:Estradiol / Q000031:analogs & derivatives / Q000494:pharmacology*; D004965:Estrogen Antagonists / Q000494:pharmacology; D000077267:Fulvestrant; D006801:Humans; D007700:Kinetics
D004965:Estrogen Antagonists; D005576:Colforsin; D000077267:Fulvestrant; D003687:Dehydroepiandrosterone; D004958:Estradiol; D019314:Dehydroepiandrosterone Sulfate; D004054:Diethylstilbestrol; D015096:3-Hydroxysteroid Dehydrogenases