Cyclic adenosine 3',5'-monophosphate induces prolactin expression in stromal cells isolated from human proliferative endometrium

Abstract

In vitro decidualization of stromal cells isolated from human proliferative endometrium and cultured in RPMI-1640 containing 2% charcoal-treated fetal bovine serum and 0.1 U/ml insulin was achieved by adding to the medium cAMP derivatives [(Bu)2cAMP (db-cAMP) and 8-bromo-cAMP] or forskolin. PRL production under these conditions was demonstrated by documenting the synthesis of PRL mRNA (approximately 1.1 kilobase), the output of immunoprecipitable [35S]methionine-labeled PRL migrating as a single 23-kilodalton band during gel electrophoresis, and the time- and concentration-dependent secretion of PRL into the medium, measured by RIA (maximal on days 4-5 using 0.5 mM db-cAMP). Medroxyprogesterone acetate (1 microM) enhanced (1.7- to 2.5-fold) the effect of db-cAMP, 8-bromo-cAMP, or forskolin on PRL production, as evaluated by Western blotting analysis. Further evidence for a participation of db-cAMP in the decidualization process was provided by its ability to induce immunocytochemically detectable heat shock protein-27, insulin-like growth factor-binding protein-1, desmin, and laminin, all compounds produced by human decidual cells, but not expressed by stromal cells. The induction of PRL by cAMP may be a key step in the process of differentiation of fibroblast-like stromal cells to the decidual phenotype, as it has been previously reported by this laboratory that, under similar culture conditions, PRL itself is capable of inducing the production of heat shock protein-27, insulin-like growth factor-binding protein-1, desmin, and laminin in stromal cells isolated from proliferative endometrium.