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Effect of Glutathione and N-Acetylcysteine on Hepatocellular Modifications Induced by 2-Acetylaminofluorene

Institute of General Physiology, Faculty of Sciences, University of Genoa, Corso Europa 26 I 16132, Genoa, Italy

Linda Scarabelli

Institute of General Physiology, Faculty of Sciences, University of Genoa, Corso Europa 26 I 16132, Genoa, Italy

Mauro Orunesu

Institute of General Physiology, Faculty of Sciences, University of Genoa, Corso Europa 26 I 16132, Genoa, Italy

The exposure of rats to a dietary regimen containing 2-acetylaminofluorene induces a sequence of hepatocellular alterations leading to the development of preneoplastic nodules. Groups of 2-acetylaminofluorenetreated rats were given glutathione or N-acetylcysteine to evaluate the effects of these different thiols on the sequence of events that originate transformed cells. It is well known that intracellular thiols protect biological macromolecules from scavenging free radicals and electrophilic compounds produced by the metabolism of chemical agents. Male Wistar rats were maintained on a feeding regimen containing 0.05% 2-acetylaminofluorene. The diet of 2 groups of 2-acetylaminofluorene-treated animals was supplemented with either 0.1% glutathione or N-acetylcysteine. The effects in the liver of the exogenously supplied thiols during 2-acety-laminofluorene treatment were assessed evaluating DNA damage, glutathione levels, activity of marker enzymes glucose-6-phosphatase, -glutamyltranspeptidase, and glutathione-S-transferase, survival rates, and development of salivary gland tumors. Our results demonstrate that the mortality due to 2-acetylaminofluorene exposure was reduced or completely abolished by thiols and that the development of salivary gland tumors was inhibited. Exogenously supplied thiols significantly reduced DNA damage as assessed by alkaline elution. At the doses employed, glutathione and N-acetylcysteine induce early stimulation of glutathione-S-transferase, had little effect on the loss of glucose-6-phosphatase activity and scanty influence on the net increase in -glutamyltranspeptidase activity.

• 1. Ahluwalia MB, Rotstein J, Tatematsu M, Roomi MW, and Farber E (1983). Failure of glutathione to prevent liver cancer development in rats initiated with diethylnitrosamine in the resistant hepatocyte model. Carcinogenesis 4:119–121.[Abstract/Free Full Text]
• 2. Anderson ME and Meister A (1983). Transport and direct utilization of -glutamylcyst(e)ine for glutathione synthesis. Proc. Natl. Acad. Sci. USA 80:707–711.[Abstract/Free Full Text]
• 3. Becker FF (1981). Recent concepts of initiation and promotion in carcinogenesis. Am. J. Pathol. 105:3–9.[Web of Science][Medline] [Order article via Infotrieve]
• 4. Bolognesi C, Cesarone CF, and Santi L (1981). Evaluation of DNA-damage by alkaline elution technique after in vivo treatment wtih aromatic amines. Carcinogenesis 2:265–268.[Abstract/Free Full Text]
• 5. Cesarone CF, Bolognesi C, and Santi L (1979). Improved microfluorometric DNA determination in biological material using 33258 Hoechst. Anal. Biochem. 100:188–197.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• 6. Cesarone CF, Bolognesi C, and Santi L (1983). DNA-damage induced in vivo in various tissues by nitro-chlorobenzene derivatives. Mutat. Res. 116:238–246.
• 7. Cesarone CF, Fugassa E, Gallo G, Voci A, and Orunesu M (1984). Influence of the culture time on DNA damage and repair in isolated rat hepatocytes exposed to nitrochlorobenzene derivatives. Mutat. Res. 131:215–222.[Web of Science][Medline] [Order article via Infotrieve]
• 8. Chen PS, Toribara TY, and Warner W (1956). Microdetermination of phosphorus. Anal. Chem. 28:1756–1758.
• 9. Cramer JW, Miller JA, and Miller EC (1960). N-hydroxylation: a new metabolic reaction observed in the rat with the carcinogen 2-acetylaminofluorene. J. Biol. Chem. 235:885–888.[Free Full Text]
• 10. De Baun JR, Miller EC, and Miller JA (1970). N-hydroxy-2-acetylaminofluorene sulfotransferase: Its probable role in carcinogenesis and in protein-(methion-S-yl) binding in rat liver. Cancer Res. 30:577–583.[Abstract/Free Full Text]
• 11. Edwards AM (1982). Regulation of -glutamyltrans-peptidase in rat hepatocyte monolayer cultures. Cancer Res. 42:1107–1115.[Abstract/Free Full Text]
• 12. Ellman GL (1959). Tissue sulphydryl groups. Arch. Biochem. Biophys. 82:70–77.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• 13. Epstein S, Ito N, Merkow LP, and Farber E (1967). The cellular analysis of liver carcinogenesis: The induction of large hyperplastic nodules in the liver with 2-fluorenylacetamide or ethionine and some aspects of their morphology and glycogen metabolism. Cancer Res. 27:1702–1711.[Abstract/Free Full Text]
• 14. Farber E (1980). The sequential analysis of liver cancer induction. Biochim. Biophys. Acta 605:149–166.[Medline] [Order article via Infotrieve]
• 15. Fleischner G, Kirsch R, Mishkin S, and Gatmaitan Z (1976). Glutathione-S-transferases and ligandin(s). In: Glutathione: Metabolism and Functions, IM Arias and WB Jakoby (eds). Raven Press, New York, pp. 175–309.
• 16. Habig WH, Pabst MJ, and Jakoby WB (1974). Glutathione-S-transferases. J. Biol. Chem. 249:7130–7139.[Abstract/Free Full Text]
• 17. Hanigan MH and Pitot HC (1985). Gamma-gluta-myltranspeptidase: Its role in hepatocarcinogenesis. Carcinogenesis 6:165–172.[Free Full Text]
• 18. Hartree EF (1972). Determination of protein: A modification of the Lowry method that gives a linear photometric response. Anal. Biochem. 48:422–427.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• 19. Hissin PJ and Hilf R (1976). A fluorometric method for determination of oxidized and reduced glutathione in tissue. Anal. Biochem. 74:214–226.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• 20. Jensen GL and Meister A (1983). Radioprotection of human lymphoid cells by exogenously supplied glutathione is mediated by -glutamyltranspeptidase. Proc. Natl. Acad. Sci. USA 80:4714–4717.[Abstract/Free Full Text]
• 21. Kadlubar FF, Miller JA, and Miller EC (1976). Hepatic metabolism of N-hydroxy-N-methyl-4-aminoazobenzene and other N-hydroxyarylamines to reactive sulfuric acid esters. Cancer Res. 36:2350–2359.[Abstract/Free Full Text]
• 22. Karmali RA (1984). Growth inhibition and prostaglandin metabolism in the R3230 AC mammary adenocarcinoma by reduced glutathione. Cancer Biochem. Biophys. 7:147–154.[Web of Science][Medline] [Order article via Infotrieve]
• 23. Ketterer B, Kadlubar FF, Flammang T, Corne T, and Enderby G (1979). Glutathione adducts of N-methyl-4-aminoazobenzene formed in vitro and by reaction of N-benzoiloxin-methyl-4-aminoazobenzene with glutathione. Chem. Biol. Interactions 25:7–21.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• 24. King CM and Phillips B (1968). Enzyme-catalyzed reactions of the carcinogens N-hydroxy-2-fluorenyl-acetamide with nucleic acid. Science 159:1351–1357.[Abstract/Free Full Text]
• 25. Kohn KW, Erickson LC, Grimek-Ewig RA, and Friedman CA (1976). Fractionation of DNA from mammalian cells by alkaline elution. Biochemistry 15:4629–4637.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• 26. Larson A, Orrenius S, Holmgren A, and Mannervik B (eds) (1983). Functions of Glutathione: Biochemical, Physiological, Toxicological and Clinical Aspects. Raven Press, New York.
• 27. Mannervik B, Guttemberg C, Jensson H, Warholm M, and Alin P (1983). Isoenzymes of glutathione-S-transferases in rat and human tissues. In: Functions of Glutathione: Biochemical, Physiological, Toxicological and Clinical Aspects, A Larson, S Orrenius, A Holmgren, and B Mannervik (eds). Raven Press, New York, pp. 75–88.
• 28. Meerman JHN, Beland FA, Ketterer B, Srai SKS, Bruins AP, and Mulder GJ (1982). Identification of glutathione conjugates formed from N-hydroxy-2-acetylaminofluorene in the rat. Chem. Biol. Interactions 39:149–168.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• 29. Meister A and Anderson ME (1983). Glutathione. Ann. Rev. Biochem. 52:711–760.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• 30. Miller EC (1978). Some current perspectives on chemical carcinogenesis in humans and experimental animals: Presidental address. Cancer Res. 38:1479–1496.[Free Full Text]
• 31. Miller EC and Miller JA (1961). N-hydroxy-2-acetylaminofluorene: A metabolite of 2-acetylaminofluorene with increased carcinogenic activity in the rat. Cancer Res. 21:816–821.
• 32. Miller EC and Miller JA (1972). Approaches to the mechanism and control of chemical carcinogenesis. In: Environment and Cancer (The University of Texas System Cancer Center 24th Annual Symposium on Fundamental Cancer Research). William and Wilkins Co., Baltimore, pp. 5–39.
• 33. Miller JA (1970). Carcinogenesis by chemicals: An overview. G.H.A. Clowes Memorial Lecture. Cancer Res. 30:559–576.[Free Full Text]
• 34. Miller JA and Miller EC (1969). The metabolic activation of carcinogenic aromatic amines and amides. Prog. Exp. Tumor Res. 11:273–301.[Web of Science][Medline] [Order article via Infotrieve]
• 35. Morgan LR, Holdiness MR, and Gillen LE (1983). N-acetylcysteine: Its bioavailability and interaction with ifosfamide metabolites. Seminars in Oncology 1(10): 56–61.
• 36. Novi AM (1981). Regression of aflatoxin B1-induced hepatocellular carcinomas by reduced glutathione. Science 212:541–542.[Abstract/Free Full Text]
• 37. Rayshell M, Ross J, and Werbin H (1983). Evidence that N-acetoxy-N-acetyl-2-aminofluorene crosslinks DNA to protein by a free radical mechanism. Carcinogenesis 5:501–507.[Web of Science]
• 38. Swanson MA (1955). Glucose-6-phosphatase from liver. In: Methods in Enzymology, Vol. II, SP Colowick and NO Kaplan (eds). Academic Press Inc., New York, pp. 541–543.
• 39. Teebor GW and Becker FF (1971). Regression and persistence of hyperplastic nodules induced by N-2-fluorenylacetamide and their relationship to hepatocarcinogenesis. Cancer Res. 31:1–3.[Abstract/Free Full Text]
• 40. Wagner G, Frenzel H, Wefers H, and Sies H (1985). Lack of effect of long-term glutathione administration on aflatoxin B1-induced hepatoma in male rats. Chem. Biol. Interactions 53:57–68.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]

Toxicologic Pathology, Vol. 14, No. 4, 445-450 (1986)
DOI: 10.1177/019262338601400410


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