Selective regulation of RNK-16 cell matrix metalloproteinases by the EP4 subtype of prostaglandin E2 receptor.
Cell surface expression of multiple structurally and functionally distinct prostaglandin E2 (PGE2) receptors (Rs), designated the EP1, EP2, EP3, and EP4 Rs, is a principal determinant of the diverse cellular effects of PGE2. The RNK-16 line of rat large granular lymphocytes, which has served as a model for natural killer cells, coexpresses a mean of 1092 EP3 Rs and EP4 Rs per cell with a mean Kd of 2.7 nM. The presence of the EP3 and EP4 Rs and the absence of the EP1 and EP2 Rs were revealed by inhibition of [3H]PGE2 binding by the EP3/EP1R agonist sulprostone, the EP3/EP2/EP4R agonist M&B 28767, and the EP2/EP4/EP3R agonist misoprostol but not by the EP1R antagonist SC-19220 or the EP2R agonist butaprost. Functional EP4 R expression was confirmed by finding that PGE2 and misoprostol, but not butaprost or sulprostone, evoked increases in the intracellular concentration of cyclic AMP ([cAMP]) in RNK-16 cells. Matrix metalloproteinase (MMP)-1 and -3 were identified by zymography and Western blots as the principal MMPs secreted by RNK-16 cells. Secretion of both MMPs by RNK-16 cells attained a maximal level after 24 h of incubation and was enhanced significantly by 10(-9) to 10(-7) M PGE2, 10(-6) M misoprostol, and 10(-4) M dibutyryl cyclic AMP, but not by the EP3R agonist sulprostone. Thus, the effect of PGE2 on RNK-16 cell MMP secretion is mediated by an EP4 R-dependent mechanism involving increases in [cAMP]i. The migration of RNK-16 cells across micropore filters, without or with a layer of Matrigel, was stimulated chemokinetically by PGE2 and misoprostol. PGE2-elicited chemokinesis of RNK-16 cells across a Matrigel model basement membrane, but not across a microfilter alone, was suppressed by the GM 6001 inhibitor of MMP activities. Stimulation of MMP activities in RNK-16 cells by the EP4R thus facilitates migration of the NK cells across vascular basement membranes.
Zeng
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Department of Medicine, University of California Medical Center, San Francisco 94143-0711, USA.
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Goetzl
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D000527:Alprostadil / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002465:Cell Movement; D003094:Collagen / Q000378:metabolism; D017364:Collagenases / Q000378:metabolism*; D000242:Cyclic AMP / Q000494:pharmacology; D012531:Dibenz(b,f)(1,4)oxazepine-10(11H)-carboxylic acid, 8-chloro-, 2-acetylhydrazide / Q000494:pharmacology; D015232:Dinoprostone / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology*; D004338:Drug Combinations; D004591:Electrophoresis, Polyacrylamide Gel; D005109:Extracellular Matrix / Q000378:metabolism*; D007694:Killer Cells, Natural / Q000201:enzymology*; D007797:Laminin / Q000378:metabolism; D020781:Matrix Metalloproteinase 1; D019278:Matrix Metalloproteinase 3; D008666:Metalloendopeptidases / Q000378:metabolism*; D016595:Misoprostol / Q000494:pharmacology; D011448:Prostaglandin Antagonists / Q000494:pharmacology; D011453:Prostaglandins / Q000378:metabolism / Q000494:pharmacology; D011509:Proteoglycans / Q000378:metabolism; D051381:Rats; D018078:Receptors, Prostaglandin E / Q000819:agonists / Q000037:antagonists & inhibitors / Q000145:classification / Q000378:metabolism*
D004338:Drug Combinations; D007797:Laminin; D011448:Prostaglandin Antagonists; D011453:Prostaglandins; D011509:Proteoglycans; D018078:Receptors, Prostaglandin E; D016595:Misoprostol; C059485:matrigel; D012531:Dibenz(b,f)(1,4)oxazepine-10(11H)-carboxylic acid, 8-chloro-, 2-acetylhydrazide; C016767:sulprostone; D003094:Collagen; D000242:Cyclic AMP; D017364:Collagenases; D008666:Metalloendopeptidases; D019278:Matrix Metalloproteinase 3; D020781:Matrix Metalloproteinase 1; D000527:Alprostadil; C048491:butaprost; D015232:Dinoprostone