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Positive and negative regulation of cell proliferation through prostaglandin receptors in NIH‐3T3 cells
Author(s) -
Watanabe Tsuyoshi,
Satoh Hiroaki,
Togoh Masako,
Taniguchi Shigeo,
Hashimoto Yoshiaki,
Kurokawa Kiyoshi
Publication year - 1996
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/(sici)1097-4652(199611)169:2<401::aid-jcp20>3.0.co;2-a
Subject(s) - receptor , microbiology and biotechnology , 3t3 cells , cell growth , prostaglandin , cell , biology , chemistry , endocrinology , biochemistry , cell culture , genetics , transfection
Among major eicosanoids and their analogs, prostaglandin (PG) F 2α > PGD 2 > PGE 1 ≥ PGE 2 > iloprost, a stable agonist of PGI 2 , dose‐dependently stimualted DNA synthesis in quiescent NIH‐3T3 cells. PGF 2α , PGD 2 , and PGE 2 , in that order, formed inositol phosphates and elevated intracellular CA 2+ ([Ca 2+ ] i ) but did not form cAMP nor inhibit forskolin‐induced cAMP formation. Iloprost, PGI 2 , and PGE 1 induced cAMP formation dose‐dependently with an ED 50 of around 10 −7 M, and PGE 2 at more than 10 −6 M did it. [ 3 H]PGF 2α and [ 3 H]PGD 2 bindings to membranes from NIH‐3T3 cells were displaced in the order of PGF 2α > PGD 2 ≥ PGE 2 , while [ 3 H]PGE 2 binding was displaced by PGE 2 > PGD 2 ≥ PGF 2α . Expression of mRNA encoding EP1 and EP4 (EP2) subtypes could be detected by reverse transcription‐ polymerase chain reaction using primers specific for EP1 and EP4 (EP2) cDNAs, but not that of EP3 subtype mRNA. The dose dependence of cAMP formation on iloprost and PGI 2 and that of [Ca 2+ ] i , elevation on PGF 2α , D 2 , and E 2 were similar to that of [ 3 H]thymidine incorporation on the corresponding agonists. Fluprostenol (1 μM), a PGF 2α receptor agonist> 17‐phenyl‐trinor‐PGE 2 (1 μM), an EP1 receptor agonist stimulated [ 3 H]thymidine incorporation, but an EP3 receptor agonist, ONO‐AP‐324, nor an EP4 (EP2) receptor agonist, 11‐deoxy‐PGE 1 (1 μM) did not. Iloprost, dibutyryl cAMP, forskolin, or cholera toxin, when applied alone, enhanced [ 3 H]thymidine incorporation, while they inhibited [ 3 H]thymidine incorporation induced by submaximal concentrations of PGF 2α or epidermal growth factor (EGF), when applied within 12 hr after agonist stimulation. These results suggest that the proliferation of NIH‐3T3 cells is stimulated by PGs via the PGF 2α receptor, EP1 subtype of PGE receptor, and the PGI 2 /PGE 1 receptor through [Ca 2+ ] i −and cAMP‐dependent pathways, and that cAMP pathway negatively cross‐talks with [Ca 2+ ] i −or receptor tyrosine kinase‐mediated DNA synthesis in a cell cycle‐dependent manner. © 1996 Wiley‐Liss, Inc.