Glucocorticoid and polyamine involvement in zinc uptake by COMMA-1D mammary epithelial cells

Abstract

The objective was to determine if a mammary cell line shows glucocorticoid stimulation of Zn uptake, and to determine whether polyamines mediate this stimulation. 65Zn uptake by COMMA-1D mouse mammary epithelial cells over a 24-h period increased significantly in cells administered 10(-7) or 10(-6) M hydrocortisone. Incorporation of 65Zn over a 1-h period was not hydrocortisone-responsive, suggesting that these incubation times represent uptake into different pools. The rate of entry into the cells over a 15-min period was significantly increased by supplementing cells with hydrocortisone with or without prolactin. Initially, cells grown in lactogenic hormone-supplemented media (10(-6) M hydrocortisone + 5 micrograms/mL ovine prolactin) had up to 65% greater 65Zn uptake over 24 h than cells in nonsupplemented growth media. 65Zn uptake from hormone media with the spermidine synthesis inhibitor methylglyoxal-bis-(guanylhydrazone) (MGBG, 10(-5)M) added was less than from growth media. Exogenous spermidine (10(-6)-10(-3)M) added to the MGBG + hormone media increased 65Zn uptake. Difluoromethylornithine (DFMO), an inhibitor of spermidine synthesis that blocks ornithine decarboxylase, caused a slight dose-dependent decrease in 65Zn uptake over the range 10(-6)-5 x 10(-3)M (p < 0.002) and tended to decrease 65Zn-uptake in lactogenic hormone-stimulated cells with 8 h of incubation, but not at other times. These data show that Zn uptake in mammary epithelial cells can be hormonally mediated by glucocorticoids and suggest that polyamines may be intracellular mediators of this effect.