In vitro corticotropin-releasing hormone (CRH) stimulation of adrenocorticotropin release from corticotroph adenoma cells: effect of prolonged exposure to CRH and its interaction with cortisol.

To examine if down-regulation of CRH-induced ACTH release occurs in corticotroph adenoma cells as well as CRH-glucocorticoid interactions in these cells, we established primary cultures of pituitary adenoma cells obtained by transphenoidal surgery from five patients with Cushing's disease. To prevent binding of glucocorticoids by serum proteins, we used a serum-free medium containing insulin, transferrin, selenium, and epidermal growth factor. The latter was found to be essential for both basal and CRH-stimulated ACTH secretion. CRH acutely stimulated, in a dose-dependent manner, ACTH release by all adenomas studied, with an IC50 of 0.5 X 10(-9) mol/L. Prolonged exposure (10 days) to a half-maximal stimulatory concentration of CRH led to continuous stimulation of ACTH secretion. A 4-day incubation with cortisol induced a dose-dependent decrease in both basal and long term CRH-stimulated ACTH release, with no difference in the IC50 (1 X 10(-8) mol/L). These data suggest that long term exposure to CRH does not desensitize corticotroph adenoma cells. Thus, it is unlikely that long-acting analogs of CRH will be useful in the treatment of Cushing's disease. ACTH secretion from corticotroph adenomas is restrained by glucocorticoids; the sensitivity of these cells to the negative effect of glucocorticoids is not modified by long term stimulation with CRH.