Effect of leukotriene D4 on mouse embryonic stem cell migration and proliferation: involvement of PI3K/Akt as well as GSK-3β/β-catenin signaling pathways.

The actual leukotriene D(4) (LTD(4)) signaling pathways that regulate cell proliferation have not been elucidated thoroughly although fatty acid and its metabolites play a key role in regulations of embryonic functions. Thus, this study investigated the response of mouse embryonic stem (ES) cells exposed to LTD(4) and elucidated the signaling pathways as well. LTD(4) increased DNA synthesis in concentration-dependent (≥10(-7) M) and time-dependent (≥12 h) manners, as determined by [(3)H] thymidine incorporation and increased cell number. LTD(4) induced the phosphorylation of signal transducer and activator of transcription-3 (STAT3) and the increase of intracellular Ca(2+) levels via cysteinyl leukotriene (CysLT) 1 and 2 receptors. LTD(4) increased Akt activation and calcineurin expression, which were blocked by STAT3 inhibitor and calcium chelators. LTD(4)-induced glycogen synthase kinase (GSK)-3β phosphorylation was decreased by LY294002, Akt inhibitor, and cyclosporine A. LTD(4) inhibited the phosphorylation of β-catenin. In addition, LTD(4)-stimulated migration through increased activation of focal adhesion kinase (FAK) and paxillin which were blocked by Akt inhibitor and cyclosporine A. LTD(4)-induced increases in protooncogene and cell cycle regulatory proteins were blocked by cyclosporine A, FAK siRNA, and β-catenin siRNA. In conclusion, LTD(4)-stimulated mouse ES cell proliferation and migration via STAT3, phosphoinositide 3-kinases (PI3K)/Akt, Ca(2+)-calcineurin, and GSK-3β/β-catenin pathway.