In pituitary-dependent hyperadrenocorticism (Cushing's disease), the disturbed regulation of ACTH secretion is associated with neoplastic transformation of corticotropic cells. As these two phenomena are almost indissolubly connected, it is of prime importance to elucidate the factor(s) that induce corticotropic cell proliferation. Here we report on the effects of hypophysiotrophic hormones and intrapituitary growth factors on the proliferation and hormone secretion of the murine corticotropic tumour cell line AtT20/D16v, as measured by DNA content, and ACTH concentration in culture media. In addition, sensitivity to the inhibitory effect of cortisol was assessed under various conditions. Corticotropin releasing hormone (CRH) and vasopressin (AVP) induced proliferation of AtT20-cells. In contrast to that caused by AVP, the CRH-induced proliferation was associated with increased ACTH secretion, which could be inhibited by cortisol. Insulin-like growth factor-I (IGF-I), epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) also stimulated the proliferation of AtT20-cells. The proliferation of AtT20-cells was significantly inhibited by cortisol in all tests. The IGF-I-induced proliferation was the least sensitive to inhibition by cortisol. The growth factors did not stimulate ACTH secretion but IGF-I differed in that it prevented the inhibition of basal ACTH secretion by cortisol. Additional experiments (Western ligand blot analysis) concerning the relative insensitivity of IGF-I induced proliferation to inhibition by cortisol revealed that IGF-I increased the concentration of a 29 kDa IGF binding protein (IGFBP) in the culture medium. The concentration of the 29 kDa IGFBP was slightly decreased by cortisol. In conclusion, the proliferation of AtT20-cells can be stimulated by the hypophysiotrophic hormones CRH and AVP and by the intrapituitary growth factors IGF-I, EGF and bFGF. Both basal and stimulated proliferation are sensitive to inhibition by cortisol, although this effect is remarkably low in the presence of IGF-I. IGF-I induced the secretion of a 29 kDa IGFBP, which might mediate the IGF-I effects by its intrinsic mitogenic properties. In addition to loss of sensitivity to endogenous glucocorticoids, high IGF-I concentrations may be a prerequisite for clonal expansion of pituitary corticotropes.
|Number of pages||7|
|Journal||Molecular and Cellular Endocrinology|
|Publication status||Published - 28 Apr 1995|
Bibliographical noteFunding Information:
This work was supported by N.W.O. (Dutch Organization for Scientific Research). Grant 900-543-066. The authors would like to thank Adri Slob, Elpetra P.M. Sprang and Jeannette Wolfswinkel for their excellent technical support and Yvonne W.E.A. Pollak and Joop Fama for preparing figures and photos.