Fibronectin is a ubiquitous extracellular matrix (ECM) protein known to play a critical role in cell adhesion. In the present study we dissected the effects of glucocorticoids and cyclic adenosine 3',5'-monophosphate (cAMP) on FN expression in cultures of cytotrophoblasts isolated from human term placentas to identify compounds which may influence uterine-placental adherence. Based on immunoassay data, relative to controls, glucocorticoid treatment (1-1000 nM) of cytotrophoblasts specifically inhibited media levels of oncofetal FN (i.e., FNs bearing an oncofetal epitope) 65-92%. Treatment of cytotrophoblasts with androgens, estrogens, and progestins (1-1000 nM) did not markedly affect onfFN expression. Corticotropin-releasing hormone (CRH) treatment (200 nM) alone had no effect on levels on onfFN. In combination experiments using 100 nM dexamethasone (DEX), 1000 nM medroxyprogesterone acetate (MPA), 10 nM estradiol (E2) and 200 nM corticotropin-releasing hormone CRH, we observed that DEX treatment also promoted approximately an 85% reduction in media levels of onfFN. This indicated that glucocorticoids profoundly suppress FN expression in the presence of high concentrations of other steroids and pregnancy-associated paracrine effectors. To examine the influence of ECM protein composition on glucocorticoid-mediated suppression of onfFN expression, cells were inoculated on untreated culture wells or on wells coated with FN, laminin, or collagen I. We observed that DEX treatment downregulated onfFN levels 70-85% under each of these conditions, suggesting that glucocorticoid effects on FN expression were not dependent on the presence of an exogenous ECM. Treatment of cytotrophoblasts with 8-bromo-cAMP resulted in a dose-dependent reduction in onfFN expression to 3% of control levels with an EC50 of 150 nM. Based on Northern blotting, treatment of cytotrophoblasts with 100 nM DEX, 1 mM 8-bromo-cAMP, or 2 nM relaxin inhibited steady state levels of FN mRNA approximately 90% relative to controls. Our results suggest that during pregnancy glucocorticoids and compounds that alter intracellular concentrations of cAMP may profoundly suppress FN expression and therefore may have dramatic effects on uterine-placental adherence.