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Toxicologic Pathology
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Journal Article

Studies on the Mechanisms by Which Tumor Promoters Stimulate the Growth of Primary Neonatal Rat Hepatocytes

Flora Romano

Tissue Culture Laboratory, Department of Human Anatomy, University of Padua, Padua, 1–35100, Venetia, Italy and Department of Dental Histology, Faculty of Medicine, University of Verona, Verona, 1–37100, Venetia, Italy

Paola G. Andreis

Tissue Culture Laboratory, Department of Human Anatomy, University of Padua, Padua, 1–35100, Venetia, Italy and Department of Dental Histology, Faculty of Medicine, University of Verona, Verona, 1–37100, Venetia, Italy

Cintia Marchesini

Tissue Culture Laboratory, Department of Human Anatomy, University of Padua, Padua, 1–35100, Venetia, Italy and Department of Dental Histology, Faculty of Medicine, University of Verona, Verona, 1–37100, Venetia, Italy

Lucia Paccagnella

Tissue Culture Laboratory, Department of Human Anatomy, University of Padua, Padua, 1–35100, Venetia, Italy and Department of Dental Histology, Faculty of Medicine, University of Verona, Verona, 1–37100, Venetia, Italy

Ubaldo Armato

Tissue Culture Laboratory, Department of Human Anatomy, University of Padua, Padua, 1–35100, Venetia, Italy and Department of Dental Histology, Faculty of Medicine, University of Verona, Verona, 1–37100, Venetia, Italy

A single exposure to a low concentration (10-10 mol/L) of several tumor promoters, namely 12-O-tetra-decanoylphorbol-13-acetate (TPA), phenobarbital (PB), nafenopin, saccharin, teleocidin, benzoyl peroxide, butylated hydroxytoluene (BHT), dichlorodiphenyltrichloroethane (DDT), lindane, clofibrate, and melittin significantly stimulated DNA synthesis of neonatal rat hepatocytes in 4-day-old primary cultures. These cultures were kept in low-calcium (0.01 mmol/L) HiWoBa2000 synthetic medium, thereby evoking a neoplastic phenotype in otherwise normal (i.e., non-initiated) cells. The simultaneous addition of a single dose of alpha-tocopherol (10-4mol/L) or selenous acid (10-5mol/L), just as that of exogenous superoxide dismutase (SOD) (4), together with each of the above agents fully suppressed the stimulation of hepatocytic DNA synthesis by the xenobiotics. Hence, these findings strengthen the view that superoxide anions (or some other oxidizing compounds) act as the common mediators of the mitogenic effects of various tumor promoters in hepatocytes. Inhibition kinetics studies, in which TPA in a single dose (10-10mol/L) was used as the paradigmatic compound together with several kinds of inhibitors of its activity showed that the early mitogenic effects of TPA, i.e., the commitment of quiescent (GO) hepatocytes and the reentry into active cycling of hepatocytes spontaneously poised at the G1/S boundary, required oxidizing compounds, arachidonate metabolism derivatives, and plasmalemmal calcium-binding sites and transmembrane calcium fluxes. Instead, a later TPAs effect, the flow into DNA synthesis of hepatocytes previously committed to cycle, was shown to be controlled by retinoid-modulable activities, by some product(s) of the lipoxygenase pathway, and again by plasmalemmal calcium-binding sites and transmembrane calcium fluxes. Such results reveal that in the neonatal rat hepatocyte the ability to answer to a single mitogenic stimulus and the metabolic pathways by which this answer is enacted depend upon the mitotic cycle setting of the hepatocytes at the moment of the experimental treatment.

  • 1. Andreis PG and Armato U (1981). Effects of epidermal growth factor/urogastrone and associated pancreatic hormones on mitotic cycle phases and proliferation kinetics of neonatal rat hepatocytes in primary culture. Endocrinology 108: 1954–1964.[Abstract/Free Full Text]
  • 2. Andreis PG, Whitfield JF, and Armato U (1981). Stimulation of DNA synthesis and mitosis of hepatocytes in primary cultures of neonatal rat liver by arachidonic acid and prostaglandins. Exp. Cell Res. 134: 265–272.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
  • 3. Armato U, Andreis PG, Draghi E, Negri E, Mengato L, and Neri G (1978). Studies on the persistence of differentiated functions in rat hepatocytes set into primary tissue culture. II. Production of specific exportable proteins and the effect of purine cyclic nucleotides: An immunofluorescent study. In Vitro 14: 838–848.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
  • 4. Armato U, Andreis PG, and Romano F (1984). Exogenous Cu,Zn-superoxide dismutase suppresses the stimulation of neonatal rat hepatocytes’ growth by tumor promoters. Carcinogenesis 5: 1547–1555.[Abstract/Free Full Text]
  • 5. Armato U, Andreis PG, and Romano F (1985). The stimulation by the tumour promoters 12-O-tetra-decanoylphorbol-13-acetate and phenobarbital of the growth of primary neonatal rat hepatocytes. Carcinogenesis 6: 811–821.[Abstract/Free Full Text]
  • 6. Armato U, Romano F, and Andreis PG (1984). The tumor promoters TPA, phenobarbital, and nafenopin, and the prostaglandins of A, E, and F series overcome the G1/S block imposed by extracellular calcium deprivation on neonatal rat hepatocytes. Prost. Leuk. Med. 13: 237–247.[CrossRef]
  • 7. Boynton AL and Whitfield JF (1980). Stimulation of DNA synthesis in calcium-deprived T51B liver cells by the tumor promoters phenobarbital, saccharin, and 12-O-tetradecanoylphorbol-13-acetate. Cancer Res. 40: 4541–4545.[Abstract/Free Full Text]
  • 8. Brennan JK, Mansky J, Roberts G, and Lichtman MA (1975). Improved methods for reducing calcium and magnesium concentrations in tissue culture medium: Application to studies of lymphoblast proliferation in vitro. In Vitro 11: 354–360.
  • 9. Bulkley GB (1983). The role of oxygen free radicals in human disease processes. Surgery 94: 407–411.[Web of Science][Medline] [Order article via Infotrieve]
  • 10. Chafouleas JC, Bolton WE, Hidaka H, Boyd AE III, and Means AR (1982). Calmodulin and the cell cycle: Involvement in regulation of cell cycle progression. Cell 28: 41–50.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
  • 11. Crutchley DJ and Maynard JR (1983). Induction of plasminogen activator by 12-O-tetradecanoylphorbol-13-acetate and calcium ionophore. Suppression by inhibitors of fatty acid lipoxygenase. Biochim. Biophys. Acta 762: 76–85.[Medline] [Order article via Infotrieve]
  • 12. Diplock AT (1984). Vitamin E, selenium and free radicals. Med. Biol. 62: 78–80.[Web of Science][Medline] [Order article via Infotrieve]
  • 13. Draghi E, Armato U, Andreis PG, and Mengato L (1980). The stimulation by epidermal growth factor (urogastrone) of the growth of neonatal rat hepatocytes in primary culture and its modulation by serum and associated pancreatic hormones. J. Cell. Physiol. 103: 129–147.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
  • 14. Emerit I and Cerutti P (1981). Tumor promoter phorbol-12-myristate-13-acetate induces chromosomal damage via indirect action. Nature 293: 144–146.[CrossRef][Medline] [Order article via Infotrieve]
  • 15. Emerit I and Cerutti P (1982). Tumor promoter phorbol-12-myristate-13-acetate induces a clastogenic factor in human lymphocytes. Proc. Natl. Acad. Sci. (USA) 79: 7509–7513.[Abstract/Free Full Text]
  • 16. Emerit I, Levy A, and Cerutti P (1983). Suppression of tumor promoter phorbolmyristate acetate-induced chromosome breakage by antioxidants and inhibitors of arachidonic acid metabolism. Mutat. Res. 110: 327–335.[Web of Science][Medline] [Order article via Infotrieve]
  • 17. Evans CH (1983). Interesting and useful biochemical properties of lanthanides. Trends Biochem. Sci. 12: 445–449.
  • 18. Fischer SM, Gleason GL, Hardin LG, Bohrman JS, and Slaga TJ (1980). Prostaglandin modulation of phorbol ester skin tumor promotion. Carcinogenesis 1: 245–248.[Abstract/Free Full Text]
  • 19. Fischer SM, Mills GD, and Slaga TJ (1982). Inhibition of mouse skin tumor promotion by several inhibitors of arachidonic acid metabolism. Carcinogenesis 3: 1243–1245.[Abstract/Free Full Text]
  • 20. Fridovich I (1976). Oxygen radicals, hydrogenperoxide, and oxygen toxicity. In: Free Radicals in Biology, Vol. 1, WA Pryor (ed). Academic Press, New York, NY, pp. 239–277.
  • 21. Friedman J and Cerutti P (1983). The induction of ornithine decarboxylase by phorbol-12-myristate 13-acetate or by serum is inhibited by antioxidants. Carcinogenesis 4: 1425–1427.[Abstract/Free Full Text]
  • 22. Goldstein BD, Witz G, Amoruso M, and Troll W (1979). Protease inhibitors antagonize the activation of polymorphonuclear leukocyte oxygen consumption. Biochem. Biophys. Res. Commun. 88: 854–860.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
  • 23. Goodwin JS, Husby G, and Williams RC Jr (1980). Prostaglandin E and cancer growth. Cancer Immunol. Immunother. 8: 3–7.[Web of Science]
  • 24. Gryglewski RJ (1979). Effects of anti-inflammatory steroids on arachidonate cascade. In: Advances in Inflammation Research, G Weissmann, B Samuelsson, and R Paoletti, (eds). Raven Press, New York, NY, pp. 505–513.
  • 25. Hamilton JA (1983). Glucocorticoids and prostaglandins inhibit the induction of macrophage DNA synthesis by macrophage growth factor and phorbol ester. J. Cell. Physiol. 115: 67–74.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
  • 26. Hammarstrom S, Lindgren JA, Marcelo C, Duell EA, Andreson TJ, and Vorhees JJ (1979). Arachidonic acid transformation in normal and psoriatic skin. J. Invest. Dermatol. 73: 180.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
  • 27. Hecker E, Fusenig NE, Kunz W, Marks F, and Thielmann HW (1982). Carcinogenesis, Vol. 7, Cocarcinogenesis and Biological Effects of Tumour Promoters. Raven Press, New York, NY.
  • 28. Hokama Y, Cripps C, Sumida K, Mookini RK, Oishi N, Kimura LH, and Kobara TY (1981). Significant increase of plasma prostaglandins in cancer patients. Res. Commun. Chem. Pathol. Pharmacol. 31: 379–382.[Web of Science][Medline] [Order article via Infotrieve]
  • 29. Kensler TW, Busch DM, and Kozumbo WS (1983). Inhibition of tumor promotion by a biomimetic superoxide dismutase. Science 221: 75–77.[Abstract/Free Full Text]
  • 30. Klein-Szanto AJP and Slaga TJ (1982). Effects of peroxides on rodent skin: Epidermal hyperplasia and tumor promotion. J. Invest. Dermatol. 79: 30–34.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
  • 31. Janis RA and Scriabine A (1983). Sites of action of Ca2+ channel inhibitors. Biochem. Pharmacol. 32: 3499–3507.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
  • 32. Janis RA and Triggle DJ (1983). New developments in Ca2+ channel antagonists. J. Med. Chem. 26: 775–785.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
  • 33. Levine L (1982). Effects of tumor promoters on arachidonic acid metabolism by cells in culture. In: Carcinogenesis, Vol. 7, Cocarcinogenesis and Biological Effects of Tumor Promoters, E Hecker, NE Fusenig, W Kunz, F Marks, and HW Thielmann, (eds). Raven Press, New York, NY, pp. 477–494.
  • 34. Levine L (1983). Inhibition of the A-23187-stimulated leukotriene and prostaglandin biosynthesis of rat basophil leukemia (RBL-1) cells by nonsteroidal anti-inflammatory drugs, antioxidants, and calcium channel blockers. Biochem. Pharmacol. 32: 3023–3026.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
  • 35. Medina D, Lane HW, and Tracey CM (1983). Selenium and mouse mammary tumorigenesis: An investigation of possible mechanisms. Cancer Res. 43: 2460–2464.[Web of Science]
  • 36. Morisaki N, Lindsay JA, Stitts JM, Zhang H, and Cornwell DG (1984). Fatty acid metabolism and cell proliferation. V. Evaluation of pathways for the generation of lipid peroxides. Lipids 19: 381–394.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
  • 37. Nakadate T, Yamamoto S, Ishii M, and Kato R (1982). Inhibition of 12-O-tetradecanoylphorbol-13-acetate-induced epidermal ornithine decarboxylase activity by phospholipase A2 inhibitors and lipoxygenase inhibitor. Cancer Res. 42: 2841–2845.[Abstract/Free Full Text]
  • 38. Nasagawa H and Little JB (1981). Factors influencing the induction of sister chromatid exchanges in mammalian cells by 12-O-tetradecanoylphorbol-13-acetate. Carcinogenesis 2: 1141–1146.[Abstract/Free Full Text]
  • 39. Oberley LW, Bize IB, Sahu SK, Leuthauser SW, and Gruber HE (1978). Superoxide dismutase activity in normal murine liver, regenerating liver, and H6 hepatoma. J. Natl. Cancer Inst. 61: 375–379.[Web of Science][Medline] [Order article via Infotrieve]
  • 40. O'Brien TG and Diamond L (1978). Ornithine decarboxylase, polyamines, and tumor promoters. In: Carcinogenesis, Vol. 2, Mechanisms of Tumor Promotion and Cocarcinogenesis, TJ Slaga, A Sivak, and RK Boutwell, (eds). Raven Press, New York, NY, pp. 273–287.
  • 41. Oyanagui Y (1976). Participation of superoxide anions at the prostaglandin phase of carragenaan footoedema. Biochem. Pharmacol. 25: 1465–1472.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
  • 42. Ozawa K, Hanaki A, and Matsuo M (1983). Reactions of superoxide ion with tocopherol and its model compounds: Correlation between the physiological activities of tocopherols and the concentrations of chromanoxyl type radicals. Biochem. Int. 6: 685–692.[Web of Science][Medline] [Order article via Infotrieve]
  • 43. Pick E, Keisari Y, Bromberg Y, Freund M, and Yakubowski A (1982). Effect of tumor promoters in immunological systems–-The macrophage as a target cell for the action of phorbol esters. In: Carcinogenesis, Vol 7, Cocarcinogenesis and Biological Effects of Tumor Promoters, E Hecker, NE Fusenig, W Kunz, F Marks, and HW Thielmann, (eds). Raven Press, New York, NY, pp. 625–635.
  • 44. Pollard M and Luckert PH (1980). Indomethacin treatment of rats with dimethylhydrazine-induced intestinal tumors. Cancer Treatment Rep. 64: 1323–1327.[Web of Science][Medline] [Order article via Infotrieve]
  • 45. Sakamoto H, Terada M, Fujiki H, Mori M, Nakayasu M, Sugimura T, and Weinstein IB (1981). Stimulation of prostaglandin production and choline turnover in HeLa cells by lyngbyatoxin A and dihydroteleocidin B. Biochem. Biophys. Res. Commun. 102: 100–107.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
  • 46. Schulte-Hermann R, Ohde G, Schuppler J, and Timmerman-Trosiener I (1981). Enhanced proliferation of putative preneoplastic cells in rat liver following treatment with the tumor promoters phenobarbital, hexachlorocyclohexane, steroid compounds, and nafenopin. Cancer Res. 41: 2556–2562.[Abstract/Free Full Text]
  • 47. Shier WT (1979). Activation of high levels of endogenous phospholipase A2 in cultured cells. Proc. Natl. Acad. Sci. (USA) 76: 195–199.[Abstract/Free Full Text]
  • 48. Sinha BK, Trush MA, and Kalyanaraman B (1983). Free radical metabolism of VP-16 cells and inhibition of anthracycline-induced lipid peroxidation. Biochem. Pharmacol. 32: 3495–3498.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
  • 49. Slaga TJ, Fischer SM, Viaje A, Berry DL, Bracken WM, LeClerc S, and Miller DR (1978). Inhibition of tumor promotion by anti-inflammatory agents: An approach to the biochemical mechanisms of promotion. In: Carcinogenesis, Vol. 2, Mechanisms of Tumor Promotion and Cocarcinogenesis, TJ Slaga, A Sivak, and RK Boutwell (eds). Raven Press, New York, NY, pp. 173–195.
  • 50. Snoek GT and Levine L (1983). Requirements for protein synthesis and calcium for stimulation of prostaglandin synthesis in cultured rat liver cells by tumor promoters. Cancer Res. 43: 4743–4746.[Abstract/Free Full Text]
  • 51. Troll W, Witz G, Goldstein B, Stone D, and Sugimura T (1982). The role of free oxygen radicals in tumor promotion and carcinogenesis. In: Carcinogenesis, Vol. 7, Cocarcinogenesis and Biological Effects of Tumor Promoters, E Hecker, NE Fusenig, W Kunz, F Marks, and HW Thielmann (eds). Raven Press, New York, NY, pp. 593–597.
  • 52. VanBelle H (1981). R24571: A potent inhibitor of calmodulin-activated enzymes. Cell Calcium 2: 483–494.[CrossRef][Web of Science]
  • 53. Van Duuren BL (1976). Tumor promoting and carcinogenic agents. In: Chemical Carcinogenesis, CE Searle (ed). American Chemical Society Monographs, Washington, DC, pp. 24–51.
  • 54. Veigl ML, Vanaman TC, and Sedwick WC (1984). Calcium and calmodulin in cell growth and transformation. Biochim. Biophys. Acta 738: 21–48.[Medline] [Order article via Infotrieve]
  • 55. Whitfield JF, Boynton AL, MacManus JP, Rixon RH, Walker PR, and Armato U (1976). The positive regulation of cell proliferation by a calcium-cyclic AMP control couplet. In: Cyclic Nucleotides and the Regulation of Cell Growth, M Abou-Sabe (ed). Dowden, Hutchinson, and Ross, Stroudsburg, PA, pp. 97–130.
  • 56. Whitfield JF, Boynton AL, MacManus JP, Sikorska M, and Tsang BK (1979). The regulation of cell proliferation by calcium and cyclic AMP. Molec. Cell. Biochem. 27: 155–179.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
  • 57. Witz G, Goldstein BD, Amoruso M, Stone DS, and Troll W (1980). Retinoid inhibition of superoxide anion radical production by polymorphonuclear leukocytes stimulated with tumor promoters. Biochem. Biophys. Res. Commun. 97: 883–888.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
  • 58. Yager JD Jr, Pariza MW, Becker JE, and Van Potter R (1975). DNA synthesis in primary cultures of parenchymal cells isolated from regenerating rat liver. In: Liver Regeneration After Experimental Injury, R Lesch and W Reutter (eds). Stratton Intercontinental Medical Book Corporation, New York, NY, pp. 148–151.
  • 59. Zuckerman AJ, Tsiquaye KN, and Fulton F (1967). Tissue culture of human embryo liver cells and the cytotoxicity of aflatoxin B1. Br. J. Exp. Pathol. 48: 20–27.[Web of Science][Medline] [Order article via Infotrieve]

Toxicologic Pathology, Vol. 14, No. 3, 375-385 (1986)
DOI: 10.1177/019262338601400315
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