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. 1985 Apr;75(4):1381–1387. doi: 10.1172/JCI111839

Human platelet-derived growth factor stimulates prostaglandin synthesis by activation and by rapid de novo synthesis of cyclooxygenase.

A J Habenicht, M Goerig, J Grulich, D Rothe, R Gronwald, U Loth, G Schettler, B Kommerell, R Ross
PMCID: PMC425468  PMID: 3921570

Abstract

Human platelet-derived growth factor (PDGF) stimulated prostaglandin (PG) E2 synthesis in the cell cycle of Swiss 3T3 cells at two distinct time intervals, with a first plateau within 10 min and a second plateau within 2-4 h after addition of PDGF. At 4 h, the concentration of PGE2 in PDGF-stimulated cultures exceeded the quiescent control cells by a factor of 10-15. Quiescent cells incubated with up to 16 microM exogenous arachidonic acid (AA) synthesized only small amounts of PGE2. In contrast, 4 h after addition of PDGF, the concentration of PGE2 synthesized from exogenous AA exceeded that in quiescent cultures by a factor of 28. The effect of PDGF stimulation on PG synthesis from exogenous AA could not be explained by growth factor-mediated increase in the cellular free AA pool as shown in experiments using [14C]AA. PDGF also stimulated synthesis of PGI2 (prostacyclin), thromboxane, and PGF2 alpha from exogenous AA. While inhibition of protein synthesis by 10 micrograms/ml cycloheximide had no effect on the early increase in PGE2 synthesis, the second increase was completely prevented. Additionally, cycloheximide treatment at 6 h after PDGF stimulation resulted in rapid decline of PGE2 synthesis from exogenous AA. Quiescent cultures pretreated with 100 microM aspirin and stimulated by PDGF thereafter recovered from cyclooxygenase inhibition within 180 min. Our results suggest that phospholipase activation and resultant AA release is not sufficient to induce the burst of PG synthesis observed in PDGF-stimulated cells. Instead, PDGF stimulates PG synthesis by direct effects on the PG-synthesizing enzyme system, one involving a protein synthesis-independent mechanism and another that requires rapid translation of cyclooxygenase.

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Selected References

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  1. Antoniades H. N., Owen A. J. Growth factors and regulation of cell growth. Annu Rev Med. 1982;33:445–463. doi: 10.1146/annurev.me.33.020182.002305. [DOI] [PubMed] [Google Scholar]
  2. Bell R. L., Kennerly D. A., Stanford N., Majerus P. W. Diglyceride lipase: a pathway for arachidonate release from human platelets. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3238–3241. doi: 10.1073/pnas.76.7.3238. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Billah M. M., Lapetina E. G., Cuatrecasas P. Phospholipase A2 activity specific for phosphatidic acid. A possible mechanism for the production of arachidonic acid in platelets. J Biol Chem. 1981 Jun 10;256(11):5399–5403. [PubMed] [Google Scholar]
  4. Bills T. K., Smith J. B., Silver M. J. Selective release of archidonic acid from the phospholipids of human platelets in response to thrombin. J Clin Invest. 1977 Jul;60(1):1–6. doi: 10.1172/JCI108745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bonser R. W., Chandrabose K. A., Cuatrecasas P. Thrombin and bradykinin modulate prostaglandin synthetase independently of phospholipase. Adv Prostaglandin Thromboxane Res. 1980;6:259–262. [PubMed] [Google Scholar]
  6. Brotherton A. F., Hoak J. C. Prostacyclin biosynthesis in cultured vascular endothelium is limited by deactivation of cyclooxygenase. J Clin Invest. 1983 Oct;72(4):1255–1261. doi: 10.1172/JCI111081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cooper J. A., Bowen-Pope D. F., Raines E., Ross R., Hunter T. Similar effects of platelet-derived growth factor and epidermal growth factor on the phosphorylation of tyrosine in cellular proteins. Cell. 1982 Nov;31(1):263–273. doi: 10.1016/0092-8674(82)90426-3. [DOI] [PubMed] [Google Scholar]
  8. Coughlin S. R., Moskowitz M. A., Zetter B. R., Antoniades H. N., Levine L. Platelet-dependent stimulation of prostacyclin synthesis by platelet-derived growth factor. Nature. 1980 Dec 11;288(5791):600–602. doi: 10.1038/288600a0. [DOI] [PubMed] [Google Scholar]
  9. Flower R. J., Blackwell G. J. Anti-inflammatory steroids induce biosynthesis of a phospholipase A2 inhibitor which prevents prostaglandin generation. Nature. 1979 Mar 29;278(5703):456–459. doi: 10.1038/278456a0. [DOI] [PubMed] [Google Scholar]
  10. Granström E., Kindahl H. Radioimmunoassay of prostaglandins and thromboxanes. Adv Prostaglandin Thromboxane Res. 1978;5:119–210. [PubMed] [Google Scholar]
  11. Habenicht A. J., Glomset J. A., Goerig M., Gronwald R., Grulich J., Loth U., Schettler G. Cell cycle-dependent changes in arachidonic acid and glycerol metabolism in Swiss 3T3 cells stimulated by platelet-derived growth factor. J Biol Chem. 1985 Feb 10;260(3):1370–1373. [PubMed] [Google Scholar]
  12. Habenicht A. J., Glomset J. A., King W. C., Nist C., Mitchell C. D., Ross R. Early changes in phosphatidylinositol and arachidonic acid metabolism in quiescent swiss 3T3 cells stimulated to divide by platelet-derived growth factor. J Biol Chem. 1981 Dec 10;256(23):12329–12335. [PubMed] [Google Scholar]
  13. Habenicht A. J., Glomset J. A., Ross R. Relation of cholesterol and mevalonic acid to the cell cycle in smooth muscle and swiss 3T3 cells stimulated to divide by platelet-derived growth factor. J Biol Chem. 1980 Jun 10;255(11):5134–5140. [PubMed] [Google Scholar]
  14. Hammarström S. Prostaglandin production by normal and transformed 3T3 fibroblasts in cell culture. Eur J Biochem. 1977 Mar 15;74(1):7–12. doi: 10.1111/j.1432-1033.1977.tb11360.x. [DOI] [PubMed] [Google Scholar]
  15. Hong S. L., Deykin D. The activation of phosphatidylinositol-hydrolyzing phospholipase A2 during prostaglandin synthesis in transformed mouse BALB/3T3 cells. J Biol Chem. 1981 May 25;256(10):5215–5219. [PubMed] [Google Scholar]
  16. Hyman B. T., Stoll L. L., Spector A. A. Prostaglandin production by 3T3-L1 cells in culture. Biochim Biophys Acta. 1982 Nov 12;713(2):375–385. doi: 10.1016/0005-2760(82)90256-9. [DOI] [PubMed] [Google Scholar]
  17. Irvine R. F. How is the level of free arachidonic acid controlled in mammalian cells? Biochem J. 1982 Apr 15;204(1):3–16. doi: 10.1042/bj2040003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Majerus P. W. Arachidonate metabolism in vascular disorders. J Clin Invest. 1983 Nov;72(5):1521–1525. doi: 10.1172/JCI111110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Majerus P. W., Prescott S. M., Hofmann S. L., Neufeld E. J., Wilson D. B. Uptake and release of arachidonate by platelets. Adv Prostaglandin Thromboxane Leukot Res. 1983;11:45–52. [PubMed] [Google Scholar]
  20. Michell R. H. Inositol lipid metabolism in dividing and differentiating cells. Cell Calcium. 1982 Oct;3(4-5):429–440. doi: 10.1016/0143-4160(82)90028-8. [DOI] [PubMed] [Google Scholar]
  21. Michell R. H. Inositol phospholipids and cell surface receptor function. Biochim Biophys Acta. 1975 Mar 25;415(1):81–47. doi: 10.1016/0304-4157(75)90017-9. [DOI] [PubMed] [Google Scholar]
  22. Okegawa T., Jonas P. E., DeSchryver K., Kawasaki A., Needleman P. Metabolic and cellular alterations underlying the exaggerated renal prostaglandin and thromboxane synthesis in ureter obstruction in rabbits. Inflammatory response involving fibroblasts and mononuclear cells. J Clin Invest. 1983 Jan;71(1):81–90. doi: 10.1172/JCI110754. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Powell W. S. Rapid extraction of arachidonic acid metabolites from biological samples using octadecylsilyl silica. Methods Enzymol. 1982;86:467–477. doi: 10.1016/0076-6879(82)86218-6. [DOI] [PubMed] [Google Scholar]
  24. Raines E. W., Ross R. Platelet-derived growth factor. I. High yield purification and evidence for multiple forms. J Biol Chem. 1982 May 10;257(9):5154–5160. [PubMed] [Google Scholar]
  25. Ross R., Glomset J. A. Atherosclerosis and the arterial smooth muscle cell: Proliferation of smooth muscle is a key event in the genesis of the lesions of atherosclerosis. Science. 1973 Jun 29;180(4093):1332–1339. doi: 10.1126/science.180.4093.1332. [DOI] [PubMed] [Google Scholar]
  26. Roth G. J., Stanford N., Majerus P. W. Acetylation of prostaglandin synthase by aspirin. Proc Natl Acad Sci U S A. 1975 Aug;72(8):3073–3076. doi: 10.1073/pnas.72.8.3073. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rozengurt E., Stroobant P., Waterfield M. D., Deuel T. F., Keehan M. Platelet-derived growth factor elicits cyclic AMP accumulation in Swiss 3T3 cells: role of prostaglandin production. Cell. 1983 Aug;34(1):265–272. doi: 10.1016/0092-8674(83)90157-5. [DOI] [PubMed] [Google Scholar]
  28. Samuelsson B., Goldyne M., Granström E., Hamberg M., Hammarström S., Malmsten C. Prostaglandins and thromboxanes. Annu Rev Biochem. 1978;47:997–1029. doi: 10.1146/annurev.bi.47.070178.005025. [DOI] [PubMed] [Google Scholar]
  29. Sawyer S. T., Cohen S. Enhancement of calcium uptake and phosphatidylinositol turnover by epidermal growth factor in A-431 cells. Biochemistry. 1981 Oct 13;20(21):6280–6286. doi: 10.1021/bi00524a057. [DOI] [PubMed] [Google Scholar]
  30. Shier W. T. Serum stimulation of phospholipase A2 and prostaglandin release in 3T3 cells is associated with platelet-derived growth-promoting activity. Proc Natl Acad Sci U S A. 1980 Jan;77(1):137–141. doi: 10.1073/pnas.77.1.137. [DOI] [PMC free article] [PubMed] [Google Scholar]

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