Cell type-specific induction of cyclin D and cyclin-dependent kinase inhibitor p27 kip1 expression by estrogen in rat endometrium

Abstract

Cyclins, cyclin-dependent kinases (CDKs) and the CDK inhibitor p27 kip1 are known to be involved in the regulation of G 1/S phase transition by estrogen in the rodent endometrium. Little is known, however, of the cell-specific location and regulation of these proteins during this process, or the way they mediate the differential effect of estrogen in the epithelium and stroma of the endometrium. Here we studied the cell-specific regulation of D-type cyclin (D 1–3), of cyclin A and E, of CDK 2 and p27 kip1 by 17β-estradiol in the endometrium of ovariectomized rats. Time-course changes in these proteins in the endometrium of ovariectomized rats were examined by immunohistochemistry at 2, 4, 8, 12, 20, 28 and 32 h after estrogen stimulation. The expression of proliferation cell nuclear antigen (PCNA) was also studied as a marker of proliferating cells. As expected from previous studies, all the proteins investigated were up-regulated by estrogen, with peak times from 8 to 32 h. The induction of cyclin D 1 is predominant in the glandular epithelium, whereas cyclin D 3 increases mainly in the luminal epithelium. The up-regulation of p27 kip1 is restricted to stromal cells with a ‘gradient-like’ expression pattern, in which the sub-epithelial (functional) layer showed stronger staining than the basal layer. The differential regulation of cyclins and p27 kip1 in the epithelium and stroma of the endometrium appear indicative of distinct actions of estrogen in different cell types in the uterus, as D-type cyclins mediate the proliferative effect of estrogen in epithelial cells while p27 kip1 might help prevent the same effect in the stroma.