title: Induction of adherent activity in mastocytoma P-815 cells by the cooperation of two prostaglandin E2 receptor subtypes, EP3 and EP4.
abstract: In this study, we investigated the role of PGE(2) in mouse mastocytoma P-815 cell adhesion to extracellular matrix proteins (ECMs) in vitro. We report that PGE(2) accelerated ProNectin F(TM) (a proteolytic fragment of fibronectin)-mediated adhesion, which was abolished by addition of the GRGDS peptide, an inhibitor of the RDG binding site of ProNectin F(TM). We show that the cAMP level and cAMP-regulated protein kinase (PKA) activity are critical mediators of this PGE(2) effect, because the cell-permeable cAMP analogue 8-Br-cAMP accelerated P-815 cell adhesion to ProNectin F(TM) and the pharmacological inhibitor of PKA, H-89, blocked PGE(2)-mediated adhesion. Consistent with mRNA expression of the G(s)-coupled EP4- and G(i)-coupled EP3-PGE receptor subtypes, P-815 cell adhesion was accelerated by treatment with a selective EP4 agonist, ONO-AE1-329, but not a selective EP1/EP3 agonist, sulprostone. However, simultaneous treatment with ONO-AE1-329 and sulprostone resulted in augmentation of both the cAMP level and cell adhesion. The augmentation of EP3-mediated cAMP synthesis was dose-dependent, without affecting the half-maximal concentration for EP4-mediated G(s)-activity, which was inhibited by a G(i) inhibitor, pertussis toxin. In conclusion, these findings suggest that PGE(2) accelerates RGD-dependent adhesion via cooperative activation between EP3 and EP4 and contributes to the recruitment of mast cells to the ECM during inflammation.
authors.affiliation: Department of Physiological Chemistry, Graduate School of Pharmaceutical Sciences, Kyoto University, Yoshida, Sakyo-ku, Kyoto 606, Japan.
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Hatae
authors.forename: Ayumi
authors.initials: A
authors.lastname: Kita
authors.forename: Satoshi
authors.initials: S
authors.lastname: Tanaka
authors.forename: Yukihiko
authors.initials: Y
authors.lastname: Sugimoto
authors.forename: Atsushi
authors.initials: A
authors.lastname: Ichikawa
mesh_terms: D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000378:metabolism; D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D002448:Cell Adhesion; D000242:Cyclic AMP / Q000378:metabolism; D015232:Dinoprostone / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005353:Fibronectins / Q000378:metabolism; D007249:Inflammation; D007546:Isoquinolines / Q000494:pharmacology; D034801:Mastocytoma / Q000378:metabolism; D051379:Mice; D011485:Protein Binding; D011494:Protein Kinases / Q000378:metabolism; D018078:Receptors, Prostaglandin E / Q000737:chemistry* / Q000378:metabolism; D058308:Receptors, Prostaglandin E, EP3 Subtype; D058309:Receptors, Prostaglandin E, EP4 Subtype; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013449:Sulfonamides; D014407:Tumor Cells, Cultured
chemical_list: D005353:Fibronectins; D007546:Isoquinolines; C545648:Ptger3 protein, mouse; C545651:Ptger4 protein, mouse; D018078:Receptors, Prostaglandin E; D058308:Receptors, Prostaglandin E, EP3 Subtype; D058309:Receptors, Prostaglandin E, EP4 Subtype; D013449:Sulfonamides; D015124:8-Bromo Cyclic Adenosine Monophosphate; C016767:sulprostone; D000242:Cyclic AMP; D011494:Protein Kinases; D000262:Adenylyl Cyclases; D015232:Dinoprostone; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide