Gastrointestinal inflammation is mediated by the pro‐inflammatory mediators interleukin‐8 (IL‐8) and prostaglandin E2 (PGE2). PGE2 binding and coupling through EP2/4 receptor subtypes on colonic epithelial cells stimulates cyclic adenosine monophosphate (cAMP) and IL‐8 production. Here we determined the mechanisms whereby PGE2 regu‐lates IL‐8 in Caco2 colonic epithelial cells and in cells over‐expressing the EP2/4 receptors (EP2S/EP4S). PGE2 coupling through EP2 activated the transcription factor inducible cAMP early repressor (ICER), whereas coupling through EP4 receptors activated the cyclic AMP‐responsive element‐binding protein (CREB). Activation of CREB in Caco2/EP2S was protein kinase A (PKA) dependent, whereas in EP4S cells, activation of CREB occurred through the PKA and phosphatidylinositol 3‐kinase pathways. Since ICER lacks the transactivation domain, it functions as a transcription repressor as opposed to CREB. PGE2 coupling through EP2/4 receptors can therefore acts in an opposing manner to either decrease (EP2) or promote IL‐8 expression by recruiting CREB‐binding protein (CBP) (EP4), which formed a multiprotein IL‐8 enhanceosome. A novel half CRE (167CRE) and a composite NFAT1‐AP1‐like site in the IL‐8 promoter participated in binding and complex formation as confirmed by mutagenesis and expression studies. These data unravel the mechanisms by which expression of IL‐8 is controlled by different signalling pathways that are activated by PGE2 but acting through different EP receptors.