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Interleukin-1ß Induces Cyclooxygenase-2 Gene Expression in Cultured Endometrial Stromal Cells

Department of Obstetrics, Gynecology, and Reproductive Sciences, Division of Reproductive Endocrinology, and the Department of Internal Medicine, Division of Hematology (K.K.W.), University of Texas Health Science Center, Houston, Texas 77030

Address all correspondence and requests for reprints to: Jaou-Chen Huang, M.D., Department of Obstetrics, Gynecology, and Reproductive Sciences, University of Texas Medical School, 6431 Fannin, MSB 3.036, Houston, Texas 77030. E-mail: jhuang{at}obg.med.uth.tmc.edu

Increasing evidence indicates that PGs may play an obligatory role in blastocyst implantation. Cyclooxygenase (also known as PGH synthase) isozymes 1 and 2 catalyze the rate-limiting steps in the biosynthesis of PGs. The ubiquitous cyclooxygenase-1 (COX-1) subserves housekeeping functions, whereas the inducible cyclooxygenase-2 (COX-2) is expressed by limited cell types and tightly controlled. Here we report the induction of COX-2 gene expression by interleukin-1ß (IL-1ß) in cultured human endometrial stromal cells.

COX-2 activity was induced by IL-1ß (1 ng/mL); conversion of exogenous arachidonic acid to PGF₂ increased from 2.6 ± 0.6 ng/well (mean ± SEM; n = 6) to 22.2 ± 5.6 ng, but was completely blocked (2.8 ± 0.7 ng/well) by NS-398, a specific COX-2 inhibitor. Undetectable in quiescent stromal cells, messenger ribonucleic acid for COX-2 was induced 30 min after IL-1ß treatment, reached a maximum at 4 h, and decreased after 15 h. Protein synthesis was not required for induction of the COX-2 gene, as it was blocked by actinomycin D but not by cycloheximide. The 70-kDa COX-2 protein was not detected in quiescent cells, became detectable 6 h after IL-1ß treatment, and remained detectable even after 15 h. IL-1ß (0.1–100 ng/mL) increased the luciferase activity in promoterless luciferase reporter containing the 900-bp 5'-flanking sequence (-891 to +9) of the COX-2 gene in a dose-dependent manner, with an ED₅₀ of 0.1–1 ng/mL.

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