The human prostanoid DP receptor stimulates mucin secretion in LS174T cells.
This study demonstrates the localization of the prostaglandin (PG)D(2) receptor (DP) within the mucous-secreting globlet cells of the human colon by in situ hybridization, which suggests a role for DP in mucous secretion. Selective high affinity ligands were used, therefore, to evaluate DP regulation of mucous secretion in LS174T human colonic adenocarcinoma cells. The expression of hDP in LS174T cells was confirmed at the mRNA level by reverse transcriptase-polymerase chain reaction, and at the protein level by radioligand binding assays and signal transduction (cyclic AMP accumulation) assays. PGD(2) and the highly selective DP-specific agonist L-644,698 ((4-(3-(3-(3-hydroxyoctyl)-4-oxo-2-thiazolidinyl) propyl) benzoic acid) (racemate)), but not PGE(2) competed for [(3)H]-PGD(2)-specific binding to LS174T cell membranes (K:(i) values of 0.4 nM and 7 nM, respectively). The DP-specific agonists PGD(2), PGJ(2), BW245C (5-(6-carboxyhexyl)-1-(3-cyclohexyl-3-hydroxypropylhydantoin)), and L-644,698 showed similar potencies in stimulating cyclic AMP accumulation (EC(50) values: 45 - 90 nM) and demonstrated the expected rank order of potency. PGE(2) also elicited cyclic AMP production in this cell line (EC(50) value: 162 nM). The activation of cyclic AMP production by PGD(2) and L-644,698, but not PGE(2), was inhibited by the selective DP antagonist BW A868C. Thus, PGD(2) and L-644,698 act through hDP in LS174T cells. PGD(2), L-644,698 and PGE(2) (an established mucin secretagogue) potently stimulated mucin secretion in LS174T cells in a concentration-dependent manner (EC(50)<50 nM). However, BW A868C effectively antagonized only the mucin secretion mediated by PGD(2) and L-644,698 and not PGE(2). These data support a role for the DP receptor in the regulation of mucous secretion.
Wright
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DH
Department of Pharmacology & Therapeutics, McGill University, McIntyre Medical Sciences Building, 3655 Promenade Sir-William-Osler, Montréal, Québec, Canada.
Ford-Hutchinson
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AW
Chadee
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Metters
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KM
D001565:Benzoates / Q000494:pharmacology; D001667:Binding, Competitive / Q000187:drug effects; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D003106:Colon / Q000187:drug effects / Q000378:metabolism; D000242:Cyclic AMP / Q000378:metabolism; D015232:Dinoprostone / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006801:Humans; D006827:Hydantoins / Q000494:pharmacology; D017403:In Situ Hybridization; D009077:Mucins / Q000187:drug effects / Q000378:metabolism*; D015230:Prostaglandin D2 / Q000378:metabolism / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011869:Radioligand Assay; D011971:Receptors, Immunologic; D011982:Receptors, Prostaglandin / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D013844:Thiazoles / Q000494:pharmacology; D053778:Thiazolidines; D014158:Transcription, Genetic; D014316:Tritium; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000378:metabolism
D001565:Benzoates; D006827:Hydantoins; C112355:L 644698; D009077:Mucins; D012333:RNA, Messenger; D011971:Receptors, Immunologic; D011982:Receptors, Prostaglandin; D013844:Thiazoles; D053778:Thiazolidines; D014316:Tritium; C058848:BW A868C; D000242:Cyclic AMP; D015232:Dinoprostone; D015230:Prostaglandin D2; C036293:prostaglandin D2 receptor