Sphingosine 1-Phosphate in Amniotic Fluid Modulates Cyclooxygenase-2 Expression in Human Amnion-derived WISH Cells

Abstract

The metabolism of arachidonic acid, in particular the generation of prostaglandins (PGs), has been proposed to play a key role in the regulation of labor. Moreover, several extracellular proteins have been reported to modulate PG synthesis in amnion cells. In this study, we found that lipid components dissolved in the amniotic fluid modulate PG synthesis in WISH human amnion cells and identified one of these components as a sphingosine 1-phosphate (S1P). WISH cells express several S1P receptors including S1P 1, S1P 2 , and S1P 3 . When WISH cells were stimulated with S1P, PGE 2 synthesis increased in a concentration-dependent manner, showing maximal activity at around 100 nM. S1P treatment also caused the up-regulation of cyclooxygenase-2 (COX-2) mRNA and protein, which was apparent within 3-12 h of stimulation. In terms of the intracellular signaling pathway of S1P-induced WISH cell activation, we found that S1P stimulated two kinds of MAPK, ERK, and p38 kinase. We examined the roles of these two MAPKs in S1P-induced COX-2 expression. S1P-induced COX-2 expression was blocked completely by PD-98059 but not by SB-203580, suggesting that ERK has a critical role in the process.

Sphingosine 1‐phosphate/sphingosine 1‐phosphate receptor 1 signaling in rheumatoid synovium: Regulation of synovial proliferation and inflammatory gene expression

“…In addition, Kim and colleagues have demonstrated that S1P regulates COX-2 expression in amnion-derived WISH cells (8). In WISH cells, COX-2 induction promoted by S1P is partially blocked by PTX (8). Our results are consistent with these reports.…”

“…Pettus et al have demonstrated that S1P mediates COX-2 induction and PGE 2 production in response to TNF␣ in murine L929 fibroblast cells and human adenocarcinoma A549 cells (9). In addition, Kim and colleagues have demonstrated that S1P regulates COX-2 expression in amnion-derived WISH cells (8). In WISH cells, COX-2 induction promoted by S1P is partially blocked by PTX (8).…”

“…Reverse transcription (RT) and complementary DNA amplification were performed with the TaKaRa RNA polymerase chain reaction (PCR) kit (Takara Shuzo, Otsu, Japan) (28). The primers used for the RT-PCR analysis have been reported previously (8). Forward and reverse primers were as follows: human S1P 1 (429-bp product) sense 5Ј-TATCAGCGCGGACAAGGAGAACAG-3Ј, antisense 5Ј-ATAGGCAGGCCACCCAGGATGAG-3Ј; human S1P 3 (394-bp product) sense 5Ј-CTGCCTGCACAATCTCCCTG-ACTG-3Ј, antisense 5Ј-GGCCCGCCGCATCTCCT-3Ј; GAPDH (246-bp product) sense 5Ј-GATGACATCAAGAA-GGTGGTGAA-3Ј, antisense 5Ј-GTCTTACTCCTTGGAG-GCCAT-GT-3Ј.…”

“…S1P affects diverse biologic processes, including cell growth, differentiation, migration, and apoptosis. Recently, S1P-mediated modulation of cyclooxygenase 2 (COX-2) expression has been reported (7)(8)(9). Importantly, S1P is involved in various pathologic conditions and has been implicated as an important mediator in angiogenesis, cancer, and autoimmunity (1).…”

Sphingosine kinase-1 mediates TNF-α-induced MCP-1 gene expression in endothelial cells: upregulation by oscillatory flow

“…It has been demonstrated that S1P activates p38 MAPK (4,14). To investigate the role of p38 MAPK in S1P-induced MCP-1 expression, HAECs were pretreated with p38 MAPK inhibitor SB-203580 (10 M) for 1 h and exposed to S1P (5 M) for 16 h. As shown in Fig.…”