Skip to main content
PMC home page
British Journal of Cancer logoLink to British Journal of Cancer
. 1988 Jul;58(1):46–51. doi: 10.1038/bjc.1988.159

Induction of peroxisome proliferation and hepatic tumours in C57BL/6N mice by ciprofibrate, a hypolipidaemic compound.

M S Rao 1, R S Dwivedi 1, V Subbarao 1, J K Reddy 1
PMCID: PMC2246483  PMID: 2901849

Abstract

The hepatic effects of ciprofibrate, a potent peroxisome proliferator, were evaluated in male C57BL/6N mice, a mouse strain with very low incidence of spontaneous liver tumour development. Dietary feeding of ciprofibrate (0.0125% or 0.025% w/w) for 2 weeks resulted in a marked proliferation of peroxisomes (9-fold increase) and several-fold increase (8- to 10-fold) in the activity of peroxisomal beta-oxidation enzymes. Feeding ciprofibrate at 0.025% concentration for 15 months followed by a 0.0125% for 6 months led to the development of hepatic adenomas in 8/14 (57%) and hepatocellular carcinomas (HCC) in 3/14 (21%) mice. In mice given 0.0125% ciprofibrate for 18 months 5 of 8 (62%) and 3 of 8 (37%) developed adenomas and HCC respectively. Similar to the findings observed in rats, both the adenomas and HCC were negative for gamma-glutamyltranspeptidase. These results in C57BL/6N mice of hepatocarcinogenic effect of ciprofibrate, a non-genotoxic chemical, indicate that peroxisome proliferation can be used as a reliable parameter to evaluate the carcinogenicity of hypolipidaemic compounds.

Full text

PDF
46

Images in this article

48Figure 1
on p.48
48Figure 2
on p.48
48Figure 3
on p.48
48Figure 4
on p.48
49Figure 5
on p.49

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Agarwal D. K., Lawrence W. H., Nunez L. J., Autian J. Mutagenicity evaluation of phthalic acid esters and metabolites in Salmonella typhimurium cultures. J Toxicol Environ Health. 1985;16(1):61–69. doi: 10.1080/15287398509530719. [DOI] [PubMed] [Google Scholar]
  2. Butterworth B. E., Bermudez E., Smith-Oliver T., Earle L., Cattley R., Martin J., Popp J. A., Strom S., Jirtle R., Michalopoulos G. Lack of genotoxic activity of di(2-ethylhexyl)phthalate (DEHP) in rat and human hepatocytes. Carcinogenesis. 1984 Oct;5(10):1329–1335. doi: 10.1093/carcin/5.10.1329. [DOI] [PubMed] [Google Scholar]
  3. Cohen A. J., Grasso P. Review of the hepatic response to hypolipidaemic drugs in rodents and assessment of its toxicological significance to man. Food Cosmet Toxicol. 1981 Oct;19(5):585–605. doi: 10.1016/0015-6264(81)90509-5. [DOI] [PubMed] [Google Scholar]
  4. Eacho P. I., Foxworthy P. S., Johnson W. D., Hoover D. M., White S. L. Hepatic peroxisomal changes induced by a tetrazole-substituted alkoxyacetophenone in rats and comparison with other species. Toxicol Appl Pharmacol. 1986 May;83(3):430–437. doi: 10.1016/0041-008x(86)90225-5. [DOI] [PubMed] [Google Scholar]
  5. Elcombe C. R., Rose M. S., Pratt I. S. Biochemical, histological, and ultrastructural changes in rat and mouse liver following the administration of trichloroethylene: possible relevance to species differences in hepatocarcinogenicity. Toxicol Appl Pharmacol. 1985 Jul;79(3):365–376. doi: 10.1016/0041-008x(85)90135-8. [DOI] [PubMed] [Google Scholar]
  6. Essigmann E. M., Newberne P. M. Enzymatic alterations in mouse hepatic nodules induced by a chlorinated hydrocarbon pesticide. Cancer Res. 1981 Jul;41(7):2823–2831. [PubMed] [Google Scholar]
  7. Fiala S., Fiala A. E., Dixon B. -Glutamyl transpeptidase in transplantable, chemically induced rat hepatomas and "spontaneous" mouse hepatomas. J Natl Cancer Inst. 1972 May;48(5):1393–1401. [PubMed] [Google Scholar]
  8. Glauert H. P., Reddy J. K., Kennan W. S., Sattler G. L., Rao V. S., Pitot H. C. Effect of hypolipidemic peroxisome proliferators on unscheduled DNA synthesis in cultured hepatocytes and on mutagenesis in Salmonella. Cancer Lett. 1984 Sep;24(2):147–156. doi: 10.1016/0304-3835(84)90130-7. [DOI] [PubMed] [Google Scholar]
  9. Goel S. K., Lalwani N. D., Fahl W. E., Reddy J. K. Lack of covalent binding of peroxisome proliferators nafenopin and Wy-14 643 to DNA in vivo and in vitro. Toxicol Lett. 1985 Jan;24(1):37–43. doi: 10.1016/0378-4274(85)90137-7. [DOI] [PubMed] [Google Scholar]
  10. Gupta R. C., Goel S. K., Earley K., Singh B., Reddy J. K. 32P-postlabeling analysis of peroxisome proliferator-DNA adduct formation in rat liver in vivo and hepatocytes in vitro. Carcinogenesis. 1985 Jun;6(6):933–936. doi: 10.1093/carcin/6.6.933. [DOI] [PubMed] [Google Scholar]
  11. Hashimoto T. Individual peroxisomal beta-oxidation enzymes. Ann N Y Acad Sci. 1982;386:5–12. doi: 10.1111/j.1749-6632.1982.tb21403.x. [DOI] [PubMed] [Google Scholar]
  12. Hess R., Stäubli W., Riess W. Nature of the hepatomegalic effect produced by ethyl-chlorophenoxy-isobutyrate in the rat. Nature. 1965 Nov 27;208(5013):856–858. doi: 10.1038/208856a0. [DOI] [PubMed] [Google Scholar]
  13. Kimbrough R. D. The carcinogenic and other chronic effects of persistent halogenated organic compounds. Ann N Y Acad Sci. 1979 May 31;320:415–418. doi: 10.1111/j.1749-6632.1979.tb56622.x. [DOI] [PubMed] [Google Scholar]
  14. Kluwe W. M., Huff J. E., Matthews H. B., Irwin R., Haseman J. K. Comparative chronic toxicities and carcinogenic potentials of 2-ethylhexyl-containing compounds in rats and mice. Carcinogenesis. 1985 Nov;6(11):1577–1583. doi: 10.1093/carcin/6.11.1577. [DOI] [PubMed] [Google Scholar]
  15. Kolarov J., Nelson B. D., Kuzela S. Rat liver protoporphyrinogen IX oxidase: site of synthesis and factor influencing its activity. Biochem Biophys Res Commun. 1983 Oct 31;116(2):383–387. doi: 10.1016/0006-291x(83)90533-8. [DOI] [PubMed] [Google Scholar]
  16. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  17. Lake B. G., Gray T. J., Foster J. R., Stubberfield C. R., Gangolli S. D. Comparative studies on di-(2-ethylhexyl) phthalate-induced hepatic peroxisome proliferation in the rat and hamster. Toxicol Appl Pharmacol. 1984 Jan;72(1):46–60. doi: 10.1016/0041-008x(84)90248-5. [DOI] [PubMed] [Google Scholar]
  18. Lalwani N. D., Alvares K., Reddy M. K., Reddy M. N., Parikh I., Reddy J. K. Peroxisome proliferator-binding protein: identification and partial characterization of nafenopin-, clofibric acid-, and ciprofibrate-binding proteins from rat liver. Proc Natl Acad Sci U S A. 1987 Aug;84(15):5242–5246. doi: 10.1073/pnas.84.15.5242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lalwani N. D., Reddy M. K., Ghosh S., Barnard S. D., Molello J. A., Reddy J. K. Induction of fatty acid beta-oxidation and peroxisome proliferation in the liver of rhesus monkeys by DL-040, a new hypolipidemic agent. Biochem Pharmacol. 1985 Oct 1;34(19):3473–3482. doi: 10.1016/0006-2952(85)90720-8. [DOI] [PubMed] [Google Scholar]
  20. Lalwani N. D., Reddy M. K., Qureshi S. A., Reddy J. K. Development of hepatocellular carcinomas and increased peroxisomal fatty acid beta-oxidation in rats fed [4-chloro-6-(2,3-xylidino)-2-pyrimidinylthio] acetic acid (Wy-14,643) in the semipurified diet. Carcinogenesis. 1981;2(7):645–650. doi: 10.1093/carcin/2.7.645. [DOI] [PubMed] [Google Scholar]
  21. Lazarow P. B. Assay of peroxisomal beta-oxidation of fatty acids. Methods Enzymol. 1981;72:315–319. doi: 10.1016/s0076-6879(81)72021-4. [DOI] [PubMed] [Google Scholar]
  22. Lazarow P. B., De Duve C. A fatty acyl-CoA oxidizing system in rat liver peroxisomes; enhancement by clofibrate, a hypolipidemic drug. Proc Natl Acad Sci U S A. 1976 Jun;73(6):2043–2046. doi: 10.1073/pnas.73.6.2043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lipsky M. M., Hinton D. E., Klaunig J. E., Goldblatt P. J., Trump B. F. Biology of hepatocellular neoplasia in the mouse. II. Sequential enzyme histochemical analysis of BALB/c mouse liver during safrole-induced carcinogenesis. J Natl Cancer Inst. 1981 Aug;67(2):377–392. [PubMed] [Google Scholar]
  24. McCann J., Choi E., Yamasaki E., Ames B. N. Detection of carcinogens as mutagens in the Salmonella/microsome test: assay of 300 chemicals. Proc Natl Acad Sci U S A. 1975 Dec;72(12):5135–5139. doi: 10.1073/pnas.72.12.5135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Miyazawa S., Furuta S., Osumi T., Hashimoto T. Turnover of enzymes of peroxisomal beta-oxidation in rat liver. Biochim Biophys Acta. 1980 Jul 3;630(3):367–374. doi: 10.1016/0304-4165(80)90285-8. [DOI] [PubMed] [Google Scholar]
  26. Moody D. E., Rao M. S., Reddy J. K. Mitogenic effect in mouse liver induced by a hypolipidemic drug, nafenopin. Virchows Arch B Cell Pathol. 1977 Apr 15;23(4):291–296. doi: 10.1007/BF02889139. [DOI] [PubMed] [Google Scholar]
  27. Moody D. E., Reddy J. K. Hepatic peroxisome (microbody) proliferation in rats fed plasticizers and related compounds. Toxicol Appl Pharmacol. 1978 Aug;45(2):497–504. doi: 10.1016/0041-008x(78)90111-4. [DOI] [PubMed] [Google Scholar]
  28. Moody D. E., Reddy J. K. Morphometric analysis of the ultrastructural changes in rat liver induced by the peroxisome proliferator SaH 42-348. J Cell Biol. 1976 Dec;71(3):768–780. doi: 10.1083/jcb.71.3.768. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ohmori T., Rice J. M., Williams G. M. Histochemical characteristics of spontaneous and chemically induced hepatocellular neoplasms in mice and the development of neoplasms with gamma-glutamyl transpeptidase activity during phenobarbital exposure. Histochem J. 1981 Jan;13(1):85–99. doi: 10.1007/BF01005842. [DOI] [PubMed] [Google Scholar]
  30. Pugh T. D., Goldfarb S. Quantitative histochemical and autoradiographic studies of hepatocarcinogenesis in rats fed 2-acetylaminofluorene followed by phenobarbital. Cancer Res. 1978 Dec;38(12):4450–4457. [PubMed] [Google Scholar]
  31. Rao M. S., Lalwani N. D., Watanabe T. K., Reddy J. K. Inhibitory effect of antioxidants ethoxyquin and 2(3)-tert-butyl-4-hydroxyanisole on hepatic tumorigenesis in rats fed ciprofibrate, a peroxisome proliferator. Cancer Res. 1984 Mar;44(3):1072–1076. [PubMed] [Google Scholar]
  32. Rao M. S., Reddy J. K. Peroxisome proliferation and hepatocarcinogenesis. Carcinogenesis. 1987 May;8(5):631–636. doi: 10.1093/carcin/8.5.631. [DOI] [PubMed] [Google Scholar]
  33. Rao M. S., Subbarao V., Reddy J. K. Peroxisome proliferator-induced hepatocarcinogenesis: histochemical analysis of ciprofibrate-induced preneoplastic and neoplastic lesions for gamma-glutamyl transpeptidase activity. J Natl Cancer Inst. 1986 Oct;77(4):951–956. [PubMed] [Google Scholar]
  34. Rao M. S., Tatematsu M., Subbarao V., Ito N., Reddy J. K. Analysis of peroxisome proliferator-induced preneoplastic and neoplastic lesions of rat liver for placental form of glutathione S-transferase and gamma-glutamyltranspeptidase. Cancer Res. 1986 Oct;46(10):5287–5290. [PubMed] [Google Scholar]
  35. Rao M. S., Usuda N., Subbarao V., Reddy J. K. Absence of gamma-glutamyl transpeptidase activity in neoplastic lesions induced in the liver of male F-344 rats by di-(2-ethylhexyl)phthalate, a peroxisome proliferator. Carcinogenesis. 1987 Sep;8(9):1347–1350. doi: 10.1093/carcin/8.9.1347. [DOI] [PubMed] [Google Scholar]
  36. Reddy J. K., Azarnoff D. L., Hignite C. E. Hypolipidaemic hepatic peroxisome proliferators form a novel class of chemical carcinogens. Nature. 1980 Jan 24;283(5745):397–398. doi: 10.1038/283397a0. [DOI] [PubMed] [Google Scholar]
  37. Reddy J. K., Goel S. K., Nemali M. R., Carrino J. J., Laffler T. G., Reddy M. K., Sperbeck S. J., Osumi T., Hashimoto T., Lalwani N. D. Transcription regulation of peroxisomal fatty acyl-CoA oxidase and enoyl-CoA hydratase/3-hydroxyacyl-CoA dehydrogenase in rat liver by peroxisome proliferators. Proc Natl Acad Sci U S A. 1986 Mar;83(6):1747–1751. doi: 10.1073/pnas.83.6.1747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Reddy J. K., Lalwani N. D., Qureshi S. A., Reddy M. K., Moehle C. M. Induction of hepatic peroxisome proliferation in nonrodent species, including primates. Am J Pathol. 1984 Jan;114(1):171–183. [PMC free article] [PubMed] [Google Scholar]
  39. Reddy J. K., Lalwani N. D., Reddy M. K., Qureshi S. A. Excessive accumulation of autofluorescent lipofuscin in the liver during hepatocarcinogenesis by methyl clofenapate and other hypolipidemic peroxisome proliferators. Cancer Res. 1982 Jan;42(1):259–266. [PubMed] [Google Scholar]
  40. Reddy J. K., Rao M. S., Azarnoff D. L., Sell S. Mitogenic and carcinogenic effects of a hypolipidemic peroxisome proliferator, [4-chloro-6-(2,3-xylidino)-2-pyrimidinylthio]acetic acid (Wy-14, 643), in rat and mouse liver. Cancer Res. 1979 Jan;39(1):152–161. [PubMed] [Google Scholar]
  41. Reddy J. K., Rao M. S. Malignant tumors in rats fed nafenopin, a hepatic peroxisome proliferator. J Natl Cancer Inst. 1977 Dec;59(6):1645–1650. doi: 10.1093/jnci/59.6.1645. [DOI] [PubMed] [Google Scholar]
  42. Reddy J. K., Rao S., Moody D. E. Hepatocellular carcinomas in acatalasemic mice treated with nafenopin, a hypolipidemic peroxisome proliferator. Cancer Res. 1976 Apr;36(4):1211–1217. [PubMed] [Google Scholar]
  43. Reddy J. K., Reddy M. K., Usman M. I., Lalwani N. D., Rao M. S. Comparison of hepatic peroxisome proliferative effect and its implication for hepatocarcinogenicity of phthalate esters, di(2-ethylhexyl) phthalate, and di(2-ethylhexyl) adipate with a hypolipidemic drug. Environ Health Perspect. 1986 Mar;65:317–327. doi: 10.1289/ehp.8665317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Reddy J. K., Scarpelli D. G., Subbarao V., Lalwani N. D. Chemical carcinogens without mutagenic activity: peroxisome proliferators as a prototype. Toxicol Pathol. 1983;11(2):172–180. doi: 10.1177/019262338301100209. [DOI] [PubMed] [Google Scholar]
  45. Reddy J. K., Warren J. R., Reddy M. K., Lalwani N. D. Hepatic and renal effects of peroxisome proliferators: biological implications. Ann N Y Acad Sci. 1982;386:81–110. doi: 10.1111/j.1749-6632.1982.tb21409.x. [DOI] [PubMed] [Google Scholar]
  46. Reznik-Schüller H. M., Lijinsky W. Morphology of early changes in liver carcinogenesis induced by methapyrilene. Arch Toxicol. 1981 Nov;49(1):79–83. doi: 10.1007/BF00352075. [DOI] [PubMed] [Google Scholar]
  47. Rutenburg A. M., Kim H., Fischbein J. W., Hanker J. S., Wasserkrug H. L., Seligman A. M. Histochemical and ultrastructural demonstration of gamma-glutamyl transpeptidase activity. J Histochem Cytochem. 1969 Aug;17(8):517–526. doi: 10.1177/17.8.517. [DOI] [PubMed] [Google Scholar]
  48. Small G. M., Burdett K., Connock M. J. A sensitive spectrophotometric assay for peroxisomal acyl-CoA oxidase. Biochem J. 1985 Apr 1;227(1):205–210. doi: 10.1042/bj2270205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Stäubli W., Hess R., Weibel E. R. Correlated morphometric and biochemical studies on the liver cell. II. Effects of phenobarbital on rat hepatocytes. J Cell Biol. 1969 Jul;42(1):92–112. doi: 10.1083/jcb.42.1.92. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Svoboda D. J., Azarnoff D. L. Tumors in male rats fed ethyl chlorophenoxyisobutyrate, a hypolipidemic drug. Cancer Res. 1979 Sep;39(9):3419–3428. [PubMed] [Google Scholar]
  51. Tomaszewski K. E., Agarwal D. K., Melnick R. L. In vitro steady-state levels of hydrogen peroxide after exposure of male F344 rats and female B6C3F1 mice to hepatic peroxisome proliferators. Carcinogenesis. 1986 Nov;7(11):1871–1876. doi: 10.1093/carcin/7.11.1871. [DOI] [PubMed] [Google Scholar]
  52. Usuda N., Reddy M. K., Hashimoto T., Rao M. S., Reddy J. K. Tissue specificity and species differences in the distribution of urate oxidase in peroxisomes. Lab Invest. 1988 Jan;58(1):100–111. [PubMed] [Google Scholar]
  53. Ward J. M. Morphology of hepatocellular neoplasms in B6C3F1 mice. Cancer Lett. 1980 Jun;9(4):319–325. doi: 10.1016/0304-3835(80)90024-5. [DOI] [PubMed] [Google Scholar]
  54. Warren J. R., Simmon V. F., Reddy J. K. Properties of hypolipidemic peroxisome proliferators in the lymphocyte [3H]thymidine and Salmonella mutagenesis assays. Cancer Res. 1980 Jan;40(1):36–41. [PubMed] [Google Scholar]
  55. Weibel E. R. Stereological principles for morphometry in electron microscopic cytology. Int Rev Cytol. 1969;26:235–302. doi: 10.1016/s0074-7696(08)61637-x. [DOI] [PubMed] [Google Scholar]
  56. Weisburger J. H., Williams G. M. Carcinogen testing: current problems and new approaches. Science. 1981 Oct 23;214(4519):401–407. doi: 10.1126/science.7291981. [DOI] [PubMed] [Google Scholar]
  57. Yamasaki H., Wilbourn J. D., Haroun L. Use of data from short-term tests in the evaluation of the carcinogenicity of environmental chemicals to humans. Prog Clin Biol Res. 1982;109:169–180. [PubMed] [Google Scholar]

Articles from British Journal of Cancer are provided here courtesy of Cancer Research UK

RESOURCES