Molecular & Cellular Toxicology

The Korean Society of Toxicogenomics and Toxicoproteomics (대한독성유전 단백체학회)
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Analysis of Gene Expression in Carcinogen-induced Acute Hepatotoxicity

  • Oh, Jung-Hwa (Toxicogenomics Team, Korea Institute of Toxicology) ;
  • Park, Han-Jin (Toxicogenomics Team, Korea Institute of Toxicology) ;
  • Lee, Eun-Hee (Toxicogenomics Team, Korea Institute of Toxicology) ;
  • Heo, Sun-Hee (Toxicogenomics Team, Korea Institute of Toxicology) ;
  • Cho, Jae-Woo (Clinical Pathology Team, Korea Institute of Toxicology) ;
  • Kim, Yong-Bum (Clinical Pathology Team, Korea Institute of Toxicology) ;
  • Yoon, Seok-Joo (Toxicogenomics Team, Korea Institute of Toxicology)
  • Published : 2009.03.31
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Abstract

The 2-year rodent carcinogenicity test involves long-term, repetitive dosing of animals that is both time consuming and expensive. Alternative approaches have been attempted using specific transgenic or knockout mice or toxicogenomics to predict carcinogenicity without conducting a 2-year rodent test. In addition, toxicogenomic analysis of carcinogen-treated animals could also enhance our understanding of molecular mechanisms and aid in the diagnosis of acute toxicity induced by carcinogens. Therefore, we investigated transcription profiles after administering the carcinogens 4,4-dimethylformamide (DMF) and 4-biphenylamine (ABP). BALB/c male mice were treated once with DMF (650 mg/kg i.p.) or ABP (120 mg/kg p.o.). Standard blood biochemistry and histological changes were observed. Gene expression profiles in the livers of mice treated with either vehicle or the carcinogens were analyzed using the Affymetrix $GeneChip^{(R)}$ assay. In all, 1,474 differentially expressed genes in DMF- or ABP-treated mice were identified as being either up- or down-regulated over 1.5-fold (P< 0.01), and these genes were analyzed using hierarchical clustering and Ingenuity Pathways Analysis. Of these, 107 genes were consistently regulated in both carcinogen-treated groups. Genes associated with cancer were upregulated (Por, S100a10, Tes, Ctcf, Ddx21, Eapp, Nel, and Pa2g4) or downregulated (Cbs and Gch1). Toxicological function analysis also identified genes involved in organ toxicity, including hepatotoxicity. These data may help to identify molecular markers for acute hepatotoxicity induced by carcinogens.

Keywords

References

  1. International Conference on Harmonizaton: guidance on testing for carcinogenicity of pharmaceuticals. Notice, Food and Drug Administration 63:8983-8986 (1998)
  2. Powell, C. L. et al. Phenotypic anchoring of acetaminophen-induced oxidative stress with gene expression profiles in rat liver. Toxicol Sci 93:213-222 (2006) https://doi.org/10.1093/toxsci/kfl030
  3. Chung, H. et al. Comprehensive analysis of differential gene expression profiles on carbon tetrachlorideinduced rat liver injury and regeneration. Toxicol Appl Pharmacol 206:27-42 (2004) https://doi.org/10.1016/j.taap.2004.11.004
  4. Oh, J. H. et al. Gene expression profiling of early renal toxicity induced by gentamicin in mice. Mol Cell Tox 2:185-192 (2006)
  5. Oh, J. H. et al. Gene expression analysis of lung injury in rats induced by exposure to MMA-SS welding fume for 30 days. Mol Cell Toxicol 3:306-313 (2007)
  6. Kramer, J. A. et al. Acute molecular markers of rodent hepatic carcinogenesis identified by transcription profiling. Chem Res Toxicol 17:463-470 (2004) https://doi.org/10.1021/tx034244j
  7. Ellinger-Ziegelbauer, H., Stuart, B., Wahle, B., Bomann, W. & Ahr, H. J. Characteristic expression profiles induced by genotoxic carcinogens in rat liver. Toxicol Sci 77:19-34 (2004) https://doi.org/10.1093/toxsci/kfh016
  8. Ellinger-Ziegelbauer, H., Stuart, B., Wahle, B., Bomann, W. & Ahr, H. J. Comparison of the expression profiles induced by genotoxic and nongenotoxic carcinogens in rat liver. Mutat Res 575:61-84 (2005) https://doi.org/10.1016/j.mrfmmm.2005.02.004
  9. Scailteur, V. & Lauwerys, R. R. Dimethylformamide (DMF) hepatotoxicity. Toxicology 43:231-238 (1987) https://doi.org/10.1016/0300-483X(87)90082-5
  10. Van den Bulcke, M., Rosseel, M. T., Wijnants, P., Buylaert, W. & Belpaire, F. M. Metabolism and hepatotoxicity of N,N-dimethylformamide, N-hydroxymethyl-N-methylformamide, and N-methylformamide in the rat. Arch Toxicol 68:291-295 (1994) https://doi.org/10.1007/s002040050071
  11. Chieli, E., Saviozzi, M., Menicagli, S., Branca, T. & Gervasi, P. G. Hepatotoxicity and P-4502E1-dependent metabolic oxidation of N,N-dimethylformamide in rats and mice. Arch Toxicol 69:165-170 (1995) https://doi.org/10.1007/s002040050153
  12. Gescher, A. Metabolism of N,N-dimethylformamide: key to the understanding of its toxicity. Chem Res Toxicol 6:245-251 (1993) https://doi.org/10.1021/tx00033a001
  13. Ducatman, A. M., Conwill, D. E. & Crawl, J. Germ cell tumors of the testicle among aircraft repairmen. J Urol 136:834-836 (1986) https://doi.org/10.1016/S0022-5347(17)45096-8
  14. Levin, S. M. et al. Testicular cancer in leather tanners exposed to dimethylformamide. Lancet 2:1153 (1987) https://doi.org/10.1016/S0140-6736(87)91587-X
  15. Ross, R. K., Jones, P. A. & Yu, M. C. Bladder cancer epidemiology and pathogenesis. Semin Oncol 23:536-545 (1996)
  16. Vineis, P. et al. Genetically based N-acetyltransferase metabolic polymorphism and low-level environmental exposure to carcinogens. Nature 369:154-156 (1994) https://doi.org/10.1038/369154a0
  17. Yu, M. C. et al. Acetylator phenotype, aminobiphenylhemoglobin adduct levels, and bladder cancer risk in white, black, and Asian men in Los Angeles, California. J Natl Cancer Inst 86:712-716 (1994) https://doi.org/10.1093/jnci/86.9.712
  18. Vineis, P. et al. DNA adducts in urothelial cells: relationship with biomarkers of exposure to arylamines and polycyclic aromatic hydrocarbons from tobacco smoke. Int J Cancer 65:314-316 (1996) https://doi.org/10.1002/(SICI)1097-0215(19960126)65:3<314::AID-IJC6>3.0.CO;2-2
  19. Harris, A. J., Dial, S. L. & Casciano, D. A. Compari-son of basal gene expression profiles and effects of hepatocarcinogens on gene expression in cultured primary human hepatocytes and HepG2 cells. Mutat Res 549:79-99 (2004) https://doi.org/10.1016/j.mrfmmm.2003.11.014
  20. Hartley, D. P. et al. Activators of the rat pregnave X receptor differentially modulate hepatic and intestinal gene expression. Mol Pharmacol 65:1159-1171 (2004) https://doi.org/10.1124/mol.65.5.1159
  21. Heijne, W. H. et al. Bromobenzene-induced hepatotoxicity at the transcriptome level. Toxicol Sci 79:411-422 (2004) https://doi.org/10.1093/toxsci/kfh128
  22. Huang, Q. et al. Gene expression profiling reveals multiple toxicity endpoints induced by hepatotoxicants. Mutat Res 549:147-167 (2004) https://doi.org/10.1016/j.mrfmmm.2003.12.020
  23. Hamadeh, H. K. et al. Gene expression analysis reveals chemical-specific profiles. Toxicol Sci 67:219-31 (2002) https://doi.org/10.1093/toxsci/67.2.219
  24. Thomas, R. S. et al. Identification of toxicologically predictive gene sets using cDNA microarrays. Mol Pharmacol 60:1189-1194 (2001) https://doi.org/10.1124/mol.60.6.1189
  25. Waring, J. F. et al. Identifying toxic mechanisms using DNA microarray: evidence that an experimental inhibitor of cell adhesion molecule expression signals through the aryl hydrocarbon nuclear receptor. Toxicology 181:537-550 (2002) https://doi.org/10.1016/S0300-483X(02)00477-8
  26. Tolando, R., Zanovello, A., Ferrara, R., Iley, J. N. & Manno, M. Inactivation of rat liver cytochrome P450 (P450) by N,N-dimethylformamide and N,N-dimethylacetamide. Toxicol Lett 124:101-111 (2001) https://doi.org/10.1016/S0378-4274(01)00384-8
  27. Wang, S. C., Chung, J. G., Chen, C. H. & Chen, S. C. 2- and 4-Aminobiphenyls induce oxidative DNA damage in human hepatoma (Hep G2) cells via different mechanisms. Mutat Res 593:9-21 (2006) https://doi.org/10.1016/j.mrfmmm.2005.06.023
  28. Roy, P., Tretyakov, O., Wright, J. & Waxman, D. J. Stereoselective metabolism of ifosfamide by human P-450s 3A4 and 2B6. Favorable metabolic properties of R-enantiomer. Drug Metab Dispos 27:1309-1318 (1999)
  29. Jakubowski, A. et al. TWEAK induces liver progenitor cell proliferation. J Clin Invest 115:2330-2340 (2005) https://doi.org/10.1172/JCI23486
  30. Drusco, A. et al. Knockout mice reveal a tumor suppressor function for Testin. Proc Natl Acad Sci U S A 102:10947-10951 (2005) https://doi.org/10.1073/pnas.0504934102
  31. Goodman & Gilman''s: The Pharmacological Basis of Therapeutics, 9th edition.
  32. Mori, Y. et al. Instabilotyping reveals unique mutational spectra in microsatellite-unstable gastric cancers. Cancer Res 62:3641-3645 (2002)
  33. Livak, K. J. & Schmittgen, T. D. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)). Method 25:402-408 (2001) https://doi.org/10.1006/meth.2001.1262