The effect of the trichloroethylene metabolites trichloroacetate and dichloroacetate on peroxisome proliferation and DNA synthesis in cultured human hepatocytes

  • J.E. Walgren
  • D.T. Kurtz
  • J.M. McMillan


Dichloroacetate (DCA) and trichloroacetate (TCA) are metabolites of the environmental contaminant trichloroethylene (TCE) that are thought to be responsible for its hepatocarcinogenicity in B6C3F1 mice. TCA and DCA induce peroxisomal proliferation and are mitogenic in rodent liver. The susceptibility of humans to TCA- and DCA-induced hepatocarcinogenesis is unknown. The current studies were aimed at using both primary and long-term human hepatocyte cultures to study the effects of TCA, DCA, and a potent peroxisome proliferator, WY-14,643, on peroxisomal activity and DNA synthesis in human hepatocytes. Peroxisome proliferation, as assessed by palmitoyl-CoA oxidation activity, was below the limit of detection in all human cell lines tested. However, the human cell lines did display small but significant increases in CYP450 4A11 levels following treatment with WY-14,643 (0.1 mmol/L), indicting that the CYP 4A11 gene may be regulated by peroxisome proliferator-activated receptor α in humans. Similarly to their effect in rodent hepatocyte cultures, TCA and DCA were not complete mitogens in human hepatocyte cultures. In fact, DNA synthesis tended to be significantly decreased following treatment of the cells with WY-14,643, TCA, or DCA. In contrast to rodent hepatocyte responses, TCA and DCA did not increase palmitoyl-CoA oxidation and caused a decrease in DNA synthesis in human hepatocyte cultures, suggesting that humans may not be susceptible to TCA- and DCA-induced hepatocarcinogenesis.

cell proliferation dichloroacetate human hepatocytes peroxisome proliferation trichloroethylene trichloroacetate 


  1. Anttila A, Pukkala E, Sallmen M, Hernberg S, Hemminki K. Cancer incidence among Finnish workers exposed to halo-genated hydrocarbons. J Occup Environ Med. 1995;37(7): 797-806.PubMedGoogle Scholar
  2. ATSDR. US Department of Health and Human Services, Agency for Toxic Substances and Disease Registry. Toxicologic profile for trichloroethylene; 1997.Google Scholar
  3. Axelson O, Selden A, Andersson K, Hogstedt C. Updated and expanded Swedish cohort study on trichloroethylene and cancer risk. J Occup Med. 1994;36:556-62.PubMedGoogle Scholar
  4. Bardot O, Clemencet MC, Malki MC, Latruffe N. Delayed effects of ciprofibrate on rat liver peroxisomal properties and proto-oncogene expression. Biochem Pharmacol. 1995;50:1001-6.PubMedCrossRefGoogle Scholar
  5. Bentley P, Calder I, Elcombe C, Grasso P, Stringer D, Wiegand HJ. Hepatic peroxisome proliferation in rodents and its significance for humans. Food Chem Toxicol. 1993;31:857-907.PubMedCrossRefGoogle Scholar
  6. Blaauboer BJ, von Holsteijn CWM, Bleumink R, et al. The effect of beclobric acid and clofibric acid on peroxisomal ?-oxidation and peroxisome proliferation in primary cultures of rat, monkey, and human hepatocytes. Biochem Pharmacol. 1990;40:521-8.PubMedCrossRefGoogle Scholar
  7. Blumcke S, Schwartzkopff W, Lobeck H, Edmondson NA, Prentice DE, Blane GF. Influence of fenofibrate on cellular and subcellular structure in hyperlipidemic patients. Atherosclerosis. 1983;46:105-16.PubMedCrossRefGoogle Scholar
  8. Buben JA, O'Flaherty EJ. Delineation of the role of metabolism in the hepatotoxicity of trichloroethylene and perchloroethylene: a dose-effect study. Toxicol Appl Pharmacol. 1985;78:105-22.PubMedCrossRefGoogle Scholar
  9. Bull RJ, Sanchez IM, Nelson MA, Larson JL, Lansing AJ. Liver tumor induction in B6C3F1 mice by dichloroacetate. Toxicology. 1990;63:341-59.PubMedCrossRefGoogle Scholar
  10. Bull RJ, Templin M, Larson JL, Stevens DK. The role of dichloroacetate in the hepatocarcinogenicity of trichloroethylene. Toxicol Lett. 1993;68:203-11.PubMedCrossRefGoogle Scholar
  11. Cherkaoui-Malki M, Lone YC, Corral-Debrinski M, Latruffe N. Differential proto-oncogene mRNA induction from rats treated with peroxisome proliferators. Biochem Biophys Res Commun. 1990;173:855-61.PubMedCrossRefGoogle Scholar
  12. Cohen AJ, Grasso P. Evidence for and possible mechanisms of non-genotoxic carcinogenesis in rodent liver. Mutat Res. 1981;248:271-90.Google Scholar
  13. DeAngelo AB, Daniel FB, Stober JA, Olson GR. The carcinogenicity of dichloroacetic acid in the male B6C3F1 mouse. Fundam Appl Toxicol. 1991;16:337-47.PubMedCrossRefGoogle Scholar
  14. DeAngelo AB, Daniel FB, Most BM, Olson GR. The carcinogenicity of dichloroacetic acid in the male Fischer 344 rat. Toxicology. 1996;114:207-21.PubMedCrossRefGoogle Scholar
  15. Dees C, Travis C. The mitogenic potential of trichloroethylene in B6C3F1 mice. Toxicol Lett. 1993;69:129-37.PubMedCrossRefGoogle Scholar
  16. Dees C, Travis C. Trichloroacetate stimulation of liver DNA synthesis in male and female mice. Toxicol Lett. 1994;70(3): 343-55.PubMedCrossRefGoogle Scholar
  17. Dekant W, Schulz A, Metzler M, Henschler D. Absorption, elimination, and metabolism of trichloroethylene: a quantitative comparison between mice and rats. Xenobiotica. 1986;16:143.PubMedCrossRefGoogle Scholar
  18. De La Iglesia FA, Lewis JE, Buchanan RA, Marcus EL, McMahon G. Light and electron microscopy of liver in hyperlipoproteinemic patients under long-term gemfibrozil treatment. Atherosclerosis. 1982;43:19-37.PubMedCrossRefGoogle Scholar
  19. Eacho PI, Lanier TL, Brodhecker CA. Hepatocellular DNA synthesis in rats given peroxisome proliferating agents: comparison of WY-14,643 to clofibric acid, nafenopin and LY171883. Carcinogenesis. 1991;12:1557-61.PubMedGoogle Scholar
  20. Elcombe CR. Species differences in carcinogenicity and peroxisome proliferation due to trichloroethylene: a biochemical human hazard assessment. Arch Toxicol Suppl. 1985;8:6-17.PubMedGoogle Scholar
  21. Elcombe CR, Mitchell A. Peroxisome proliferation due to di(2-ethyhexyl)phthalate: species differences and possible mechanisms. Environ Health Perspect. 1986;70:211-9.PubMedGoogle Scholar
  22. Everhart JL, Kurtz DT, McMillan JM. Dichloroacetic acid induction of peroxisome proliferation in cultured hepatocytes. J Biochem Mol Toxicol. 1998;12(6):351-9.PubMedCrossRefGoogle Scholar
  23. Gariot P, Barrat E, Drouin P, et al. Morphometric study of human hepatic cell modifications induced by fenofibrate. Metabolism. 1987;36(3):203-10.PubMedCrossRefGoogle Scholar
  24. Gerbracht U, BurschW, Kraus P, et al. Effects of the hypolipidemic drugs nafenopin and clofibrate on phenotypic expression and cell death (apoptosis) in altered foci of rat liver. Carcinogenesis. 1990;11:617-24.PubMedGoogle Scholar
  25. Gervois P, Torra IP, Chinetti G, et al. A truncated human peroxisome proliferator-activated receptor a splice variant with dominant negative activity. Mol Endocrinol. 1999; 13(9):1535-49.PubMedCrossRefGoogle Scholar
  26. Gibson-D'Ambrosio RE, Crowe DL, Shuler CE, D'Ambrosio SM. The establishment and continuous subculturing of normal human adult hepatocytes: expression of differentiated liver functions. Cell Biol Toxicol. 1993;9:385-403.PubMedCrossRefGoogle Scholar
  27. Goll V, Alexandre E, Viollon-Abadie C, Nicod L, Jaeck D, Richert L. Comparison of the effects of various peroxisome proliferators on peroxisomal enzyme activities, DNA synthesis, and apoptosis in rat and human hepatocyte cultures. Toxicol Appl Pharmacol. 1999;160:21-32.PubMedCrossRefGoogle Scholar
  28. Gonzalez FJ, Peters JM, Cattley RC. Mechanism of action of the nongenotoxic peroxisome proliferators: role of the peroxisome proliferator-activated receptor ?. J Natl Cancer Inst. 1998;90(22):1702-9.PubMedCrossRefGoogle Scholar
  29. Hardell L, Eriksson M, Lenner P, Lundgren E. Malignant lymphoma and exposure to chemicals, especially organic solvents, chlorophenols and phenoxy acids: a case-control study. Br J Cancer. 1981;43:169-76.PubMedGoogle Scholar
  30. Hasmall SC, James NH, Soames AR, Roberts RA. The peroxisome proliferator nafenopin does not suppress hepatocyte apoptosis in guinea pig liver in vivo nor in human hepatocytes in vitro. Arch Toxicol. 1998;72:777-83.PubMedCrossRefGoogle Scholar
  31. Hawkins JM, Jones WE, Bonner FW, Gibson GG. The effect of peroxisome proliferators on microsomal, peroxisomal, and mitochondrial enzyme activities in the liver and kidney. Drug Metab Rev. 1988;18:441-515.Google Scholar
  32. Herren-Freund SL, Pereira MA, Khoury MD, Olson G. The carcinogenicity of trichloroethylene and its metabolites, trichloroacetic acid and dichloroacetic acid, in mouse liver. Toxicol Appl Pharmacol. 1987;90:183-9.PubMedCrossRefGoogle Scholar
  33. Issemann I, Prince R, Tugwood J, Green S. A role for fatty acids and liver fatty acid binding protein in peroxisome proliferation? Biochem Soc Trans. 1992;20:824-7.PubMedGoogle Scholar
  34. James NH, Roberts RA. Species differences in response to peroxisome proliferators correlate in vitro with induction of DNA synthesis rather than suppression of apoptosis. Carcinogenesis. 1996;17(8):1623-32.PubMedGoogle Scholar
  35. Karam WG, Ghanayem BI. Induction of replicative DNA synthesis and PPAR?-dependent gene transcription by Wy-14,643 in primary rat hepatocyte and non-parenchymal cell co-cultures. Carcinogenesis. 1997;18(11):2077-83.PubMedCrossRefGoogle Scholar
  36. Kraupp-Grasl B, Huber W, Taper H, Schulte-Hermann R. Increased susceptibility of aged rats to hepatocarcinogenesis by the peroxisome proliferator nafenopin and the possible involvement of altered liver foci occurring spontaneously. Cancer Res. 1991;51:666-71.PubMedGoogle Scholar
  37. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227:680-5.PubMedCrossRefGoogle Scholar
  38. Lazarow PB. Assay of peroxisomal beta-oxidation of fatty acids. Methods Enzymol. 1981;72:315-9.PubMedCrossRefGoogle Scholar
  39. Ledwith BJ, Manam S, Troilo P, Joslyn DJ, Galloway SM, Nichols WW. Activation of immediate-early gene expression by peroxisome proliferators in vitro. Mol Carcinog. 1993;8:20-7.PubMedGoogle Scholar
  40. Maltoni C, Lefemine G, Cotti G, Perino G. Long-term carcinogenicity bioassays on trichloroethylene administered by inhalation to Sprague-Dawley rats and Swiss and B6C3F1 mice. Ann NY Acad Sci. 1988;534:316-42.PubMedGoogle Scholar
  41. Marsman DS, Cattley RC, Conway JG, Popp JA. Relationship of hepatic peroxisome proliferation and replicative DNA synthesis to the hepatocarcinogenicity of the peroxisome proliferators di(2-ethylhexyl)phthalate and [4-chloro-6-(2,3-xylidino)-2-pyrimidinylthio]acetic acid (WY-14,643) in rats. Cancer Res. 1988;48:6739-44.PubMedGoogle Scholar
  42. Michalopoulos GK. Liver regeneration: molecular mechanisms of growth control. FASEB J. 1990;4(2):176-87.PubMedGoogle Scholar
  43. Muerhoff AS, Griffin KJ, Johnson EF. The peroxisome proliferator-activated receptor mediates the induction of CYP4A6, a cytochrome P450 fatty acid omega-hydrolase, by clofibric acid. J Biol Chem. 1992;267:19051-3.PubMedGoogle Scholar
  44. NTP (National Toxicology Program). Carcinogenesis studies of trichloroethylene in F344/N rats and B6C3F1 mice. NTP Technical Report; no. 243; 1990.Google Scholar
  45. Palmer CN, Hsu MH, Griffin KJ, Raucy JL, Johnson EF. Peroxisome proliferator activated receptor-alpha expression in human liver. Mol Pharmacol. 1998;53(1):14-22.PubMedGoogle Scholar
  46. Parzefall W, Erber E, Sedivy R, Schulte-Hermann R. Testing for induction of DNA synthesis in human hepatocyte primary cultures by rat liver tumor promoters. Cancer Res. 1991;51:1143-7.PubMedGoogle Scholar
  47. Peters JM, Cattley RC, Gonzalez FJ. Role of PPAR? in the mechanism of action of the nongenotoxic carcinogen and peroxisome proliferator Wy-14,643. Carcinogenesis. 1997; 18(11):2029-33.PubMedCrossRefGoogle Scholar
  48. Pereira MA. Carcinogenic activity of dichloroacetic acid and trichloroacetic acid in the liver of female B6C3F1 mice. Fundam Appl Toxicol. 1996;31:192-9.PubMedCrossRefGoogle Scholar
  49. Perrone CE, Williams GM. Rodent hepatocarcinogenic peroxi-some proliferators induce proliferation of rat hepatocytes in primary mixed cultures with rat liver epithelial cells. Cancer Lett. 1998;123:27-33.PubMedCrossRefGoogle Scholar
  50. Perrone CE, Shao L, Williams GM. Effect of hepatocarcinogenic peroxisome proliferators on fatty acyl-CoA oxidase, 272 DNA synthesis, and apoptosis in cultured human and rat hepatocytes. Toxicol Appl Pharmacol. 1998;150:277-86.PubMedCrossRefGoogle Scholar
  51. Plant NJ, Horley N, Savory RL, Elcombe CR, Gray TJ, Bell DR. The peroxisome proliferators are hepatocyte mitogens in chemically-definedmedia: glucocorticoid-induced PPAR? is linked to peroxisome proliferator mitogenesis. Carcinogenesis. 1998;19(5):925-31.PubMedCrossRefGoogle Scholar
  52. Reddy JK, Rao MS. Oxidative DNA damage caused by persistent peroxisome proliferation: its role in hepatocarcinogenesis. Mutat Res. 1989;214:63-8.PubMedGoogle Scholar
  53. Richmond RE, Carter JH, Carter HW, Daniel FB, DeAngelo AB. Immunohistochemical analysis of dichloroacetic acid (DCA)-induced hepatocarcinogenesis in male Fischer (F344) rats. Cancer Lett. 1995;92:67-76.PubMedCrossRefGoogle Scholar
  54. Rouisse L, Chakrabarti SK. Dose-dependent metabolism of trichloroethylene and its relevance to hepatotoxicity in rats. Environ Res. 1986;40:450-8.PubMedCrossRefGoogle Scholar
  55. Smith PK, Krohn RI, Hermanson GT, et al. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985;150:76-85. [Published erratum appears in Anal Biochem. 1987 (May 15); 163(1):279].PubMedCrossRefGoogle Scholar
  56. Spirtas R, Stewart PA, Lee JS, et al. Retrospective cohort mortality study of workers at an aircraft maintenance facility. I. Epidemiological results. Br J Ind Med. 1991;48: 515-30.PubMedGoogle Scholar
  57. Stauber AJ, Bull RJ. Differences in phenotype and cell replicative behavior of hepatic tumors induced by dichloroacetic acid (DCA) and trichloroacetic acid (TCA). Toxicol Appl Pharmacol. 1997;144:235-46.PubMedCrossRefGoogle Scholar
  58. Styles JA, Wyatt I, Coutts C. Trichloroacetic acid: studies on uptake and effects on hepatic DNA and liver growth in mouse. Carcinogenesis. 1991;12(9):1715-9.PubMedGoogle Scholar
  59. Uden PC, Miller JW. Chlorinated acids and chloral in drinking water. J AmWaterWorks Assoc. 1983;75:524-7.Google Scholar
  60. Walgren JE, McMillan JM, Kurtz DT. Trichloroacetate and dichloroacetate are not complete mitogens in primary rat hepatocyte cultures. [Manuscript in review.]Google Scholar
  61. Woodyatt NJ, Lambe KG, Myers KA, Tugwood JD, Roberts RA. The peroxisome proliferator (PP) response element upstream of the human acyl-CoA oxidase gene is inactive among a sample human population: significance for species differences in response to PP's. Carcinogenesis. 1999;20(3): 369-72.PubMedCrossRefGoogle Scholar
  62. Wong O, Morgan R. Final report: historical prospective mortality study of aircraft employees at Air Force plant No. 44 Almeda, CA (USA). ENSR Health Sciences; 1990.Google Scholar
  63. Vanden Heuvel JP. Peroxisome proliferator-activated receptors (PPARs) and carcinogenesis. Toxicol Sci. 1999;47:1-8.PubMedCrossRefGoogle Scholar

Copyright information

© Kluwer Academic Publishers 2000

Authors and Affiliations

  • J.E. Walgren
    • 1
  • D.T. Kurtz
    • 1
  • J.M. McMillan
    • 1
  1. 1.Cell and Molecular Pharmacology and Experimental TherapeuticsMedical University of South CarolinaCharlestonUSA

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