Tumor-Promoting Effects of 2,3,7,8-Tetrachlorodibenzo- p -Dioxin and Phenobarbital in Initiated Weanling Sprague-Dawley Rats: A Quantitative,Phenotypic,and ras p21 Protein Study
Free accessResearch articleFirst published online May, 1994
Tumor-Promoting Effects of 2,3,7,8-Tetrachlorodibenzo- p -Dioxin and Phenobarbital in Initiated Weanling Sprague-Dawley Rats: A Quantitative,Phenotypic,and ras p21 Protein Study
In an initiation-promotion protocol, female weanling Sprague-Dawley rats were initiated with 10 mg/kg nitrosodiethylamine and promotion was started after 30 days. Promotion regimens were as follows: 2,3,7,8-tetrachlorodibenzo-para-dioxin (TCDD; 150 ppt in diet) continuously until day 450; phenobarbital (PB; 500 ppm in diet) until day 170; PB until day 170, followed by TCDD until day 240; and PB until day 170, followed by a basal diet (BD) until day 240 and subsequently TCDD from days 240 to 450. TCDD fed to initiated rats had a promoting effect on the development of adenosine triphosphatase-negative altered hepatocellular foci (AHF). At 450 days, the volume fraction of liver occupied by AHF was increased in initiated rats given TCDD continuously and in those given PB followed by TCDD, whereas the mean volume of AHF was significantly larger in initiated rats given TCDD continuously. PB and TCDD promoted similar phenotypes of AHF as seen in hemotoxylin and eosin-stained sections, but the eosinophilic phenotype most closely correlated with the development of hepatocellular neoplasms. The protooncogene product ras p21 protein was present in the majority of PB- and TCDD-promoted AHF, hepatocellular adenomas, and hepatocellular carcinomas. Eosinophilic AHF and ras p21 protein expression most closely correlated with neoplastic development, suggesting that these cell populations, when promoted, may be at greater risks for developing into neoplasms.
1. Bannasch P (1988). Phenotypic cellular changes as indicators of stages during neoplastic development. In: Theories of Carcinogenesis, OH Iversen (ed). Hemisphere Publishing Corporation, Washington, D.C., pp. 231–249.
2.
2. Bannasch P, Zerban H, and Hacker HJ (1985). Foci of altered hepatocytes, rat. In: Monographs on Pathology of Laboratory Animals—Digestive System, TC Jones, U Mohr, and RD Hunt (eds). Springer-Verlag, New York, pp. 10–30.
3.
3. Barbacid M (1987). ras genes. Ann. Rev. Biochem.56: 779–827.
4.
4. Bos JL (1988). The ras gene family and human carcinogenesis. Mutat. Res.195: 255–271.
5.
5. Boume JA (1983). Handbook of Immunoperoxidase Staining Methods.DAKO Corporation, Santa Barbara, California, pp. 1–37.
6.
6. Campbell HA, Pitot HC, Potter VR, and Laishes BA (1982). Application of quantitative stereology to the evaluation of enzyme altered foci in rat liver. Cancer Res.42: 465–472.
7.
7. Chandar N, Lombardi B, Schultz W, and Locker J (1987). Analysis of ras genes and linked viral sequences in rat hepatocarcinogenesis. AJP129: 232–241.
8.
8. Dragan YP and Pitot HC (1992). Commentary. The role of the stages of initiation and promotion in phenotypic diversity during hepatocarcinogenesis in the rat. Carcinogenesis13: 739–750.
9.
9. Evarts RP, Nagy P, Nakatsukasa H, Marsden E, and Thorgeirsson SS (1989). In vivo differentiation of rat liver oval cells into hepatocytes. Cancer Res.49: 1541–1547.
10.
10. Fausto N (1990). Oval cells and liver carcinogenesis: An analysis of cell lineages in hepatic tumors using oncogene transfection techniques. Prog. Clin. Biol. Res.331: 325–334.
11.
11. Fiorucci G and Hall A (1988). All three human ras genes are expressed in a wide range of tissues. Biochim. Biophys. Acta950: 81–83.
12.
12. Galand P, Jacobovitz D, and Alexandre K (1988). Immunohistochemical detection of c-Ha-ras oncogene p21 product in pre-neoplastic and neoplastic lesions during hepatocarcinogenesis in rats. Int. J. Cancer41: 155–161.
13.
13. Goldsworthy T, Campbell HA, and Pitot HC (1984). The natural history and dose-response characteristics of enzyme-altered foci in rat liver following phenobarbital and diethylnitrosamine administration. Carcinogenesis5: 67–71.
14.
14. Goyette M, Dolan M, Kaufmann W, Kaufman D, Shank PR, and Fausto N (1988). Transforming activity of DNA from rat liver tumors induced by the carcinogen methyl(acetoxy-methyl)nitrosamine. Molec. Carcinogen.1: 26–32.
15.
15. Harada T, Maronpot RR, Morris RW, Stitzel KA, and Boorman GA (1989). Morphological and sterological characterization of hepatic foci of cellular alteration in control Fischer 344 rats. Toxicol. Pathol.17: 579–593.
16.
16. Ishikawa T, Takayama S, and Kitagawa T (1980). Correlation between time of partial hepatectomy after a single treatment with diethylnitrosamine and induction of adenosinetriphosphatase-deficient islands in rat liver. Cancer Res.40: 4261–4264.
17.
17. Keenan RE, Paustenbach DJ, Wenning RJ, Parsons AH (1991). Pathology reevaluation of the Kociba et al. (1978) bioassay of 2,3,7,8-TCDD: Implications for risk assessment. J. Toxicol. Environ. Health34: 279–296.
18.
18. Kociba RJ, Keyes DG, Beyer JE, Carreon RM, Wade CE, Dittenber DA, Kalnins RP, Frauson LE, Park CN, Barnard SD, Hummel RA, and Humiston CG (1978). Results of a two-year chronic toxicity and oncogenicity study of 2,3,7,8-tetrachlorodibenzo-p-dioxin in rats. Toxicology and Applied Pharmacology46: 279–303.
19.
19. Lucier GW, Tritscher A, Goldsworthy T, Foley J, Clark G, Goldstein J, and Maronpot R (1991). Ovarian hormones enhance 2,3,7,8-tetrachlorodibenzo-p-dioxin-mediated increases in cell proliferation and preneoplastic foci in a two-stage model for rat hepatocarcinogenesis. Cancer Res.51: 1391–1397.
20.
20. Makino R, Hayashi K, Sato S, and Sugimura T (1984). Expressions of the c-Ha-ras and c-myc genes in rat liver tumors. Biochem. Biophys. Res. Commun.119: 1096–1102.
21.
21. Maronpot RR, Montgomery CA Jr, Boorman GA, and McConnell EE (1986). National Toxicology Program nomenclature for hepatoproliferative lesions of rats. Toxicol. Pathol.14: 263–273.
22.
22. McMahon G, Hanson L, Lee JJ, and Wogan GN (1986). Identification of an activated c-Ki-ras oncogene in rat liver tumors induced by anatoxin B1. Proc. Natl. Acad. Sci. USA83: 9418–9422.
23.
23. National Toxicology Program (1982). Carcinogenesis Bioassay of 2,3,7,8-Tetrachlorodibenzo-p-dioxin in Osborne-Mendel Rats and B6C3F1 Mice (Gavage Study). Technical Report 209, Carcinogenesis Testing Program, Research Triangle Park, North Carolina.
24.
24. Pitot HC, Barsness L, Goldsworthy T, and Kitagawa T (1978). Biochemical characterization of stages of hepatocarcinogenesis after a single dose of diethylnitrosamine. Nature (London)271: 456–458.
25.
25. Pitot HC, Goldsworthy T, Campbell HA, and Poland A (1980). Quantitative evaluation of the promotion by 2,3,7,8-tetrachlorodibenzo-p-dioxin of hepatocarcinogenesis. Cancer Res.40: 3616–3620.
26.
26. Poland A and Knutson JC (1982). 2,3,7,8-Tetrachlorodibenzo-p-dioxin and related halogenated aromatic hydrocarbons: Examination of the mechanism of toxicity. Annu. Rev. Pharmacol. Toxicol.22: 517–554.
27.
27. Rumsby PC, Barrass NC, Phillimore HE, and Evans JG (1991). Analysis of the Ha-ras oncogene in C3H/He mouse liver tumours derived spontaneously or induced with diethylnitrosamine or phenobarbitone. Carcinogenesis12: 2331–2336.
28.
28. Schulte-Hermann R, Kraupp-Grasl B, Bursch W, Gerbracht U, and Timmermann-Trosiener I (1989). Effects of non-genotoxic hepatocarcinogens phenobarbital and nafenopin on phenotype and growth of different populations of altered foci in rat liver. Toxicol. Pathol.17: 642–650.
29.
29. Stowers SJ, Wiseman RW, Ward JW, Miller EC, Miller JA, Anderson MW, and Eva A (1988). Detection of activated proto-oncogenes in N-nitrosodiethylamine-induced liver tumors: A comparison between B6C3F1 mice and Fischer 344 rats. Carcinogenesis9: 271–276.
30.
30. Strom SC and Faust JB (1990). Oncogene activation and hepatocarcinogenesis. Pathobiology58: 153–167.
31.
31. Tatematsu M, Tsuda H, Shirai T, Masui T, and Ito N (1987). Placental glutathione S-transferase (GST-3P) as a new marker for hepatocarcinogenesis: In vivo short-term screening for hepatocarcinogens. Toxicol. Pathol.15: 60–68.
33. Wachstein M and Meisel E (1966). Microscopic histochemical methods for the demonstration of enzymes. In: Selected Histochemical and Histopathological Methods, SW Thompson and RD Hunt (eds). Charles C Thomas Publisher, Springfield, Illinois, pp. 647–652.
34.
34. Ward JM, Pardue RL, Junker JL, Takahashi K, Shih TY, and Weislow OS (1986). Immunocytochemical localization of RasHa p21 in normal and neoplastic cells in fixed tissue sections from Harvey sarcoma virus-infected mice. Carcinogenesis7: 645–651.
35.
35. Ward JM, Perantoni AO, and Santos E (1989). Comparative immunohistochemical reactivity of monoclonal and polyclonal antibodies to H-ras p21 in normal and neoplastic tissues of rodents and humans. Oncogene4: 203–213.
36.
36. Yaswen P, Goyette M, Shank PR, and Fausto N (1985). Expression of c-Ki-ras, c-Ha-ras, and c-myc in specific cell types during hepatocarcinogenesis. Mol. Cell Biol.5: 780–786.
37.
37. Zabik MJ, Kaczmar SW, and D'Itry FM (1985). Parts per trillion residues of 2,3,7,8-tetrachlorodibenzo-p-dioxin in Michigan fish. In: Chlorinated Dioxins and Dibenzofurans in the Total Environment, LH Keith, C Rappe, and G Choudhary (eds). Butterworth Publishers, Boston, p. 103.
