This study examines the regulation of major enzymes in prostaglandin E₂ (PGE₂) synthesis by glucocorticoids in separate cultures of human amnion epithelial and fibroblast cells at term. Cytosolic phospholipase A₂ (cPLA₂), cytosolic PGES (cPGES), and microsomal PGES (mPGES) mRNA were expressed at similar levels in both cell types, whereas a greater prostaglandin H synthase type 2 (PGHS-2) mRNA expression was observed in amnion fibroblasts than in epithelial cells. Amnion fibroblasts produced 50-fold more PGE₂ per cell than epithelial cells. Dexamethasone (0.01–1 μm) increased PGE₂ production in amnion fibroblasts in a concentration-dependent manner but did not affect PGE₂ production in amnion epithelial cells. Both mRNA and protein expression of cPLA₂ and PGHS-2 but not cPGES and mPGES were increased in a dose-dependent manner by dexamethasone (0.01–1 μm) in amnion fibroblasts. Induction of cPLA₂ and PGHS-2 mRNA by dexamethasone was blocked by RU486. Dexamethasone did not affect PGHS-2, cPGES, and mPGES mRNA expression in amnion epithelial cells. In conclusion, amnion fibroblasts express a higher level of PGHS-2 mRNA and produced more PGE₂ per cell than amnion epithelial cells at term of human pregnancy. Glucocorticoids increase PGE₂ production only in the amnion fibroblasts mainly through induction of cPLA₂ and PGHS-2 expression.