Sex steroids in synergy with prostaglandins (PG) are involved in the regulation of cyclic ovarian function. In this study, we investigated the mRNA expression of three genes involved in arachidonic acid (AA) metabolism and hence PG production in domestic cats: PG‐endoperoxide synthase (PTGS2), PGF_2α synthase (PGFS) and PGE₂ synthase (PGES). Feline endometria (n = 16) were collected at oestrus and mid and late phases of pseudopregnancy. In addition, the effects of E₂ and/or P₄ on PG secretion and gene expression on endometrial explants were studied in an in vitro culture system. Expression levels of all examined genes were up‐regulated at the mid phase of pseudopregnancy. The effects of E₂ and/or P₄ treatment on both PG secretion and expression of the genes were observed after 12 h of culture. Expression of PGES was significantly up‐regulated by E₂ plus P₄ at oestrus and the mid phase of pseudopregnancy and was also up‐regulated by a single treatment with P₄ at late pseudopregnancy (p < 0.05). Simultaneous incubation with E₂ and P₄ up‐regulated PTGS2 gene expression at oestrus and mid‐luteal phase (p < 0.05). Progesterone plus E₂ significantly increased PGE₂ secretion at oestrus and the mid phase of pseudopregnancy. However, treatment with E₂ and/or P₄ affected neither PGF_2α secretion nor PGFS expression at any phase after 12 h of culture. The overall findings indicate that genes involved in PG synthesis are up‐regulated at the mid phase of pseudopregnancy. An increase in PGE₂ secretion and up‐regulation of PGES and PTGS2 are the main responses of the endometrium to treatment with E₂ and P₄ at oestrus and the mid phase of pseudopregnancy in the cat. These data support the hypothesis that ovarian sex steroids via endometrial PGE₂ are involved in endocrine homoeostasis, especially at oestrus and the mid, but not the late, phase of pseudopregnancy in cats.