Summary
In order to search for possible mediators involved in the transient radiomimetic effectiveness of TPA and related compounds early changes in the AA metabolism of HeLa cells prelabeled with 1-14C-AA have been analyzed. Maximum release of AA with different concentrations of TPA (3×10-9 to 3×10-5 M) was observed after 2–3 h treatment in the presence of 10% calf serum. Released AA was reincorporated by the cells after that period, a phenomenon which was largely abolished or delayed by cycloheximide. Reincorporation of released AA was observed in the presence of 10% fresh serum as well as with 0.5% BSA, and appears to be due to an induction of responsible enzyme(s) by the phorbol ester. The earliest metabolites of AA produced via the cyclooxygenase such as PGE2 and PGF2α and via lipoxygenases such as 12-, and 15-hydroxyeicosatetraenoic acids appear in small amounts and after later time points. AA release exhibited a pluriphasic dose response to TPA with maxima at 3×10-8 M and ≧10-5 M. Comparative dose response measurements with respect to AA release were established using various promoting skin mitogens which exhibited the following order of potency: TPA > teleocidin ≈ RPA > mezerein ≫ EPA>4-O-Me-TPA. For reasons discussed it appears unlikely that AA, Prostaglandins, or hydroxyeicosatetraenoic acid products play a significant role as mediators of the radiomimetic effects of TPA in G2 of the cell cycle.
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Abbreviations
- AA:
-
arachidonic acid
- BSA:
-
bovine serum albumin
- EPA:
-
12-O-ethacrynylphorbol-13-acetate
- 4-O-Me-TPA:
-
4-O-methyl-TPA
- PGE:
-
prostaglandin E2
- RPA:
-
12-O-retinoylphorbol-13-acetate
- TPA:
-
12-O-tetradecanoylphorbol-13-acetate
References
Anderson KE, Whitlon DS, Mueller GC (1985) Role of fatty acid structure in the reversible activation of phosphatidylcholine synthesis in lymphocytes. Biochim Biophys Acta 835:360–368
Ballou LR, Cheung WY (1985) Inhibition of human platelet phospholipase A2 activity by unsaturated fatty acids. Proc Natl Acad Sci USA 82:371–375
Billah MM, Lapetina EG, Cuatrecasas P (1981) Phospholipase A2 activity specific for phosphatidic acid. J Biol Chem 256:5399–5403
Blackwell GJ, Carnuccio R, Di Rosa M, Flower RJ, Ivanyi J, Langham CS, Parente L, Persico P, Wood J (1983) Suppression of arachidonate oxidation by glucocorticoid-induced antiphospholipase peptides. In: Samuelsson B, Paoletti R, Ramwell P (eds) Advances in prostaglandin, thromboxane, and leucotriene research. Raven Press, New York, 11:65–71
Carnuccio R, Di Rosa M, Persico P (1980) Hydrocortisone-induced inhibitor of prostaglandin biosynthesis in rat leucocytes. Br J Pharmacol 68:14–16
Carnuccio R, Di Rosa M, Flower RJ, Pinto A (1981) The inhibition by hydrocortisone of prostaglandin biosynthesis in rat peritoneal leucocytes is correlated with intracellular macrocortin levels. Br J Pharmacol 74:322–324
Cornwell DG, Morisaki N (1984) Fatty acid paradoxes in the control of cell proliferation, and cooxidation reactions. In: Pryor WA (ed) Free radicals in biology. Academic Press, New York, 6:95–148
Crutchley DJ, Conanan LB, Maynard JR (1980) Induction of plasminogen activator and prostaglandin biosynthesis in HeLa cells by TPA. Cancer Res 40:849–852
Fischer SM (1985) Arachidonic acid metabolism and tumor promotion. In: Fischer SM, Slaga TJ (eds) Arachidonic acid metabolism and tumor promotion. Martinus Nijhoff, Boston, pp21–47
Flower RJ, Blackwell GJ (1976) The importance of phospholipase A2 in prostaglandin biosynthesis. Biochem Pharmacol 25:285–291
Flower RJ, Blackwell GJ (1979) Anti-inflammatory steroids induce biosynthesis of a phospholipase A2 inhibitor which prevents prostaglandin generation. Nature 278:456–459
Flower RJ, Wood JN, Parente L (1984) Macrocortin and the mechanism of action of glucocorticoids. In: Otterness J, Capetola R, Wong S (eds) Advances in inflammation research. Raven Press, New York, 7:61–70
Fürstenberger G, Marks F (1985) Prostaglandins, epidermal hyperplasia and skin tumor promotion. In: Fischer SM, Slaga TJ (eds) Arachidonic acid metabolism and tumor promotion. Martinus Nijhoff, Boston, pp49–72
Fürstenberger G, Berry DL, Sorg B, Marks F (1981) Skin tumor promotion by phorbol esters is a two-stage process. Proc Natl Acad Sci USA 78:7722–7726
Fürstenberger G, Sorg B, Marks F (1983) Tumor promotion by phorbol esters in skin: evidence for a memory effect. Science 220:89–91
Fürstenberger G, Kinzel V, Schwarz M, Marks F (1985) Partial inversion of the initiation-promotion sequence of multistage tumorigenesis in the skin of NMRI mice. Science 230:76–78
Hecker E (1971) Isolation and characterization of the cocarcinogenic principles from croton oil. Methods Cancer Res 6:439–484
Hirata F, Matusda K, Notsu Y, Hattori T, Carmine R (1984) Phosphorylation at a tyrosine residue of lipomodulin in mitogenstimulated murine thymocytes. Proc Natl Acad Sci USA 81: 4717–4721
Hirata F, Schiffmann E, Venkatasubramania K, Salomon D, Axelrod J (1980) A phospholipase A2 inhibitory protein in rabbit neutrophils induced by glucocorticoids. Proc Natl Acad Sci USA 77:2533–2536
Irvine RF (1982) How is the level of free arachidonic acid controlled in mammalian cells? Biochem J 204:3–16
Isakson PC, Raz A, Denny SE, Wyche A, Needleman P (1977) Hormonal stimulation of arachidonate release from isolated perfused organs. Relationship to prostaglandin biosynthesis. Prostaglandins 14:853–871
Kaszkin M (1986) Einfluß von Glucocorticoiden auf die durch Tumorpromotor TPA induzierte Freisetzung von Arachidonsäure und Prostaglandin E2 aus HeLa-Zellen. Diplomarbeit Universität Heidelberg
Kinzel V, Kreibich G, Hecker E, Suess R (1979) Stimulation of choline incorporation in cell cultures by phorbol derivatives and its correlation with their irritant and tumor-promoting activity. Cancer Res 39:2743–2750
Kinzel V, Richards J, Stöhr M (1980) Tumor promoter TPA mimics irradiation effects on the cell cycle of HeLa cells. Science 210:429–431
Kinzel V, Richards J, Stöhr M (1981) Early effects of the tumor-promoting phorbol ester 12-O-tetradecanoylphorbol-13-acetate on the cell cycle. Cancer Res 41:300–305
Kinzel V, Loehrke H, Goerttler K, Fürstenberger G, Marks F (1984a) Suppression of the first stage of phorbol 12-tetradecanoate 13-acetate-effected tumor promotion in mouse skin by nontoxic inhibition of DNA synthesis. Proc Natl Acad Sci USA 81:5858–5862
Kinzel V, Richards J, Marks F, Fürstenberger F (1984b) Radio mimetic activity of phorbol esters exerted in HeLa cells in comparison with their tumor-promoting capacity. Cancer Res 44:139–143
Kinzel V, Fürstenberger G, Loehrke H, Marks F (1986) Three-stage tumorigenesis in mouse skin: DNA synthesis as a prerequisite for the conversion stage induced prior to inhibition. Carcinogenesis 7:779–782
Levine L (1981) Arachidonic acid transformation and tumor production. Adv Cancer Res 35:49–79
Levine L (1982) Effects of tumor promoters on arachidonic acid metabolism by cells in culture. In: Hecker E, Fusenig NE, Kunz W, Marks F, Thielmann HW (eds) Carcinogenesis VII: Cocarcinogenesis and biological effects of tumor promoters. Raven Press, New York, pp 477–494
Levine L, Hassid A (1977) Epidermal growth factor stimulates prostaglandin biosynthesis by canine kidney (MDCK) cells. Biochem Biophys Res Commun 76:1181–1187
Levine L, Pong S, Hong SL, Tam S (1977) Deacylation of phospholipids: prerequisite for prostaglandin synthesis by transformed fibroblasts. In: Kharasch N, Fried J (eds) Biochemical aspects of prostaglandins and thromboxanes. Proceedings of the 1976 Intra-Science Research Foundation Symposium Santa Monica, California, Academic Press, New York, pp 15–38
Little JB, Nagasawa H, Weichselbaum RR, Lovett DR, Schmit A, Kochenburger RJ, Blumberg PM (1985) Differing patterns of cytotoxicity of the phorbol ester 12-O-tetradecanoylphorbol-13-acetate in various human cell strains. Carcinogenesis 6:1703–1708
Mufson RA, DeFeo D, Weinstein JB (1979) Effects of phorbol ester tumor promoters on arachidonic acid metabolism in chick embryo fibroblasts. Mol Pharmacol 16:569–578
Ohuchi K, Watanabe M, Yoshizawa K, Tsurufuji S, Fujiki H, Suganuma T, Sugimura T, Levine L (1985) Stimulation of prostaglandin E2 production by TPA-type and non-TPA-type tumor promoters in macrophages and its inhibition by cycloheximide. Biochim Biophys Acta 834:42–47
Pelech SL, Cook HW, Paddon HB, Vance DE (1984) Membranenbound CTP: phosphocholine cytidylyl transferase regulates the rate of phosphatidylcholine synthesis in HeLa cells treated with unsaturated fatty acids. Biochim Biophys Acta 795:433–440
Pepinsky RB, Sinclair LK, Browning JL, Mattaliano RJ, Smart JE, Chow EP, Falbel T, Ribolini A, Garwin JL, Wallner BP (1986) Purification and partial sequence analyses of 37-kDa protein that inhibits phospholipase A2 activity from rat peritoneal exudates. J Biol Chem 261:4239–4246
Russo-Marie F, Duval D (1982) Dexamethasone-induced inhibition of prostaglandin production does not result from a direct action on phospholipase activities but is mediated through a steroid-inducible factor. Biochim Biophys Acta 712:177–185
Sakamoto H, Terada M, Fujiki H, Mori M, Nakayasu M, Sugimura T, Weinstein JB (1981) Stimulation of prostaglandin production and choline turnover in HeLa cells by lyngbyatoxin A and dihydroteleocidin B. Biochem Biophys Res Commun 102:100–107
Schildknecht H, Edelmann G, Maurer H (1970) Die Struktur des Mezereins aus der Frucht des Seidelbastes Daphne mezereum. Chemiker Zeitung 94:849
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Dedicated to Professor Dr. E. Hecker on the occasion of his 60th birthday
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Espe, U., Fürstenberger, G., Marks, F. et al. Early changes in the arachidonic acid metabolism of HeLa cells in response to the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) and related compounds. J Cancer Res Clin Oncol 113, 137–144 (1987). https://doi.org/10.1007/BF00391435
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DOI: https://doi.org/10.1007/BF00391435