The Regulation of Cyclooxygenase-2 Expressionby Interleukin-1beta in WISH cells / 대한산부인과학회잡지

OBJECTIVE: To determine of the regulation of cyclooxygenase-2 (COX-2) expression by Interleukin-1beta in WISH cells. METHODS: Amnion WISH cells were incubated in media containing increasing concentrations of IL-1beta or with various inhibitors. Increased COX-2 expression was determined by Western blot analysis with anti-COX-2 antibody. Concomitant measurements of culture media PGE2 were made by an enzyme immunoassay. RESULTS: The COX-2 and prostaglandin E2 production induced by IL-1beta increased in a dose- and time-dependent manner. One of the regulating factors that induced COX-2 by IL-1beta was protein kinase C (PKC). PKC inhibitor, Ro 31-8220 was pretreated and continued treating by IL-1beta. Then, PKC inhibitor completely blocked COX-2 protein induction by IL-1beta. In contrast, COX-2 induction by IL-1beta after pretreating PKC stimulator, phobol 12-myristate 13-acetate was potentiated with synergism. Another factor in controlling COX-2 protein induction was identified as phosphatidylinositol 3-kinase (PI 3K). COX-2 protein induction by IL-1beta after pretreating PI 3K inhibitors, wortmannin and LY294002 strongly increased. This kind of result reflected that PI 3K act as negative regulator. COX-2 induction by IL-1beta was known to be regulated in not only transcription step, but also translation step after performing experiment of actinomycin and cycloheximide treatment. CONCLUSION: COX-2 protein and prostaglandin E2 production induced by IL-1beta were controlled by many factors in amnion cell. Among those factors, PKC and PI 3K have an important role, but their control mechanism act as positive and negative, respectively.

Increased Expression of Prostaglandin H synthase by Aspirin in Cultured Cells from Amnionic Cell Line WISH Cells / 대한산부인과학회잡지

Prostaglandins which are produced from amnionic cells are known to play a major role in uterine contraction and cervical dilatation in human. Recently it is reported that aspirin increases the expression of PGHS-1 and PGHS-2 in trophoblast cells from placenta. We examined here the changes of immunoreactive prostaglandin H synthase (PGHS) level by aspirin in cultured cells from amniotic fluid and human amnionic WISH cells. PMA (10-7 M), an activator of protein kinase C increased the induction of PGHS-2 in both cells with or without fetal calf serum. PGHS-2 protein was also increased significantly by 10-4 M aspirin at 6 hours in both cells in the presence of serum but it was not increased in the absence of serum. The expression of PGHS-1 protein was enhanced by asprin but not by PMA in the absence of serum. Other anti-inflammatory drugs such as acetaminophen, indomethacin, dexame- thasone, and mefenamic acid increased the PGHS-2 protein level in WISH cells. PMA-induced PGHS-2 expression in WISH cells was not decreased by aspirin, on the contrary, the level was increased additively. Our results show that the increased expression of PGHS in amnion cells or other amniotic fluid cells by aspirin and other several anti-inflammatory drugs is through an unidentified effect rather than feedback effect by depletion of prostaglandin.