Identification of carcinogens by a selected panel of DNA damage response associated genes
Regina Stöber11Leibniz Research Centre for Working Environment and Human Factors at TU Dortmund (IfADo), Ardeystrasse 67, 44139 Dortmund, Germany
EXCLI J 2015;14:Doc1294
Recently, Bettina Maria Fischer and colleagues of the Institute of Toxicology in Karlsruhe have published data on a gene expression based predictive test system for chemical carcinogens (Fischer et al., 2015). The authors used bronchial epithelial cell lines and analyzed the expression of 95 genes by a high-throughput RT-qPCR system. The analyzed genes cover the biological motifs DNA damage response, genomic stability, cell cycle control, apoptosis and mitotic signaling (Fischer et al., 2015). In a case study using the carcinogenic compound cadmium, the authors demonstrate that genes involved in the DNA damage response were up-, while DNA repair genes were down-regulated, thereby giving a clear-cut positive result in the low micromolar concentration range.
Identification of chemical carcinogens represents a cutting-edge topic in toxicology (Liu et al., 2015; Ustündag et al., 2014; Bustaffa et al., 2014; Bach et al., 2014; Seiler et al., 2001; Westphal et al., 2000; Venkov et al., 2000). Since it is not possible to test all chemicals in long-term carcinogenicity rodent studies, fast but nevertheless accurate predictive tests are urgently needed (Zhang et al., 2015; Ireno et al., 2014; Kumar and Dhawan, 2013; Mohiuddin et al., 2014; Bertini et al., 2000; Ostby et al., 1997). Gene array studies have been frequently applied to characterize the impact of chemicals on genome-wide expression patterns in an unbiased manner (Cunningham, 2001; Ellinger-Ziegelbauer et al., 2008; Nie et al., 2006; Krug et al., 2013; Rempel et al., 2015; Lohr et al., 2015; Campos et al., 2014; Balmer et al., 2014). However, it has also been suggested that smaller subsets of genes may be sufficient for characterizing expression responses to chemicals due to the redundancy of highly correlated gene clusters (Waisberg et al., 2003; Grinberg et al., 2014; Godoy et al., 2015, 2013). This offers the advantage to use quantitative RT-PCR techniques instead of more cost intensive array based or sequencing approaches (Zellmer et al., 2010). In this context the test system presented by Fischer et al. (2015) represents an attractive approach. Of course validation studies will have to be performed, including sufficiently high numbers of positive and negative control compounds also in comparison to the already established bacterial and mammalian mutagenicity tests.
1. Bach J, Sampayo-Reyes A, Marcos R, Hernández A. Ogg1 genetic background determines the genotoxic potential of environmentally relevant arsenic exposures. Arch Toxicol. 2014;88:585-96.
2. Balmer NV, Klima S, Rempel E, Ivanova VN, Kolde R, Weng MK, et al. From transient transcriptome responses to disturbed neurodevelopment: role of histone acetylation and methylation as epigenetic switch between reversible and irreversible drug effects. Arch Toxicol. 2014;88:1451-68.
3. Bertini S, Del Carratore R, Giorgi M, Bronzetti G, Della Croce C. Genotoxic and mono-oxygenase system effects of the fungicide maneb. Arch Toxicol. 2000;74:415-20.
4. Bustaffa E, Stoccoro A, Bianchi F, Migliore L. Genotoxic and epigenetic mechanisms in arsenic carcinogenicity. Arch Toxicol. 2014;88:1043-67.
5. Campos G, Schmidt-Heck W, Ghallab A, Rochlitz K, Pütter L, Medinas DB, et al. The transcription factor CHOP, a central component of the transcriptional regulatory network induced upon CCl4 intoxication in mouse liver, is not a critical mediator of hepatotoxicity.Arch Toxicol. 2014;88:1267-80.
6. Cunningham MJ. Genomics and proteomics: the new millennium of drug discovery and development. J Pharmacol Toxicol Methods. 2000;44:291-300. Erratum in: J Pharmacol Toxicol Methods. 2001;45:85.
7. Ellinger-Ziegelbauer H, Gmuender H, Bandenburg A, Ahr HJ. Prediction of a carcinogenic potential of rat hepatocarcinogens using toxicogenomics analysis of short-term in vivo studies. Mutat Res. 2008;637:23-39.
8. Fischer BM, Neumann D, Piberger AL, Risnes SF, Köberle B, Hartwig A. Use of high-throughput RT-qPCR to assess modulations of gene expression profiles related to genomic stability and interactions by cadmium. Arch Toxicol. 2015 Nov 2. [Epub ahead of print].
9. Godoy P, Hewitt NJ, Albrecht U, Andersen ME, Ansari N, Bhattacharya S, et al. Recent advances in 2D and 3D in vitro systems using primary hepatocytes, alternative hepatocyte sources and non-parenchymal liver cells and their use in investigating mechanisms of hepatotoxicity, cell signaling and ADME. Arch Toxicol. 2013;87:1315-530.
10. Godoy P, Schmidt-Heck W, Natarajan K, Lucendo-Villarin B, Szkolnicka D, Asplund A, et al. Gene networks and transcription factor motifs defining the differentiation of stem cells into hepatocyte-like cells. J Hepatol. 2015;63:934-42.
11. Grinberg M, Stöber RM, Edlund K, Rempel E, Godoy P, Reif R, et al. Toxicogenomics directory of chemically exposed human hepatocytes. Arch Toxicol. 2014;88:2261-87.
12. Ireno IC, Baumann C, Stöber R, Hengstler JG, Wiesmüller L. Fluorescence-based recombination assay for sensitive and specific detection of genotoxic carcinogens in human cells. Arch Toxicol. 2014;88:1141-59.
13. Krug AK, Balmer NV, Matt F, Schönenberger F, Merhof D, Leist M. Evaluation of a human neurite growth assay as specific screen for developmental neurotoxicants. Arch Toxicol. 2013;87:2215-31.
14. Kumar A, Dhawan A. Genotoxic and carcinogenic potential of engineered nanoparticles: an update. Arch Toxicol. 2013;87:1883-900.
15. Liu L, Chen L, Floehr T, Xiao H, Bluhm K, Hollert H, et al. Assessment of the mutagenicity of sediments from yangtze river estuary using salmonella typhimurium/microsome assay. PLoS One. 2015;10:e0143522.
16. Lohr M, Hellwig B, Edlund K, Mattsson JS, Botling J, Schmidt M, et al. Identification of sample annotation errors in gene expression datasets. Arch Toxicol. 2015;89:2265-72.
17. Mohiuddin, Keka IS, Evans TJ, Hirota K, Shimizu H, Kono K, et al. A novel genotoxicity assay of carbon nanotubes using functional macrophage receptor with collagenous structure (MARCO)-expressing chicken B lymphocytes. Arch Toxicol. 2014;88:145-60.
18. Nie AY, McMillian M, Parker JB, Leone A, Bryant S, Yieh L, et al. Predictive toxicogenomics approaches reveal underlying molecular mechanisms of nongenotoxic carcinogenicity. Mol Carcinog. 2006;45:914-33.
19. Ostby L, Engen S, Melbye A, Eide I. Mutagenicity testing of organic extracts of diesel exhaust particles after fractionation and recombination. Arch Toxicol. 1997;71:314-9.
20. Rempel E, Hoelting L, Waldmann T, Balmer NV, Schildknecht S, Grinberg M, et al. A transcriptome-based classifier to identify developmental toxicants by stem cell testing: design, validation and optimization for histone deacetylase inhibitors. Arch Toxicol. 2015;89:1599-618.
21. Seiler F, Rehn B, Rehn S, Bruch J. Evidence of a no-effect level in silica-induced rat lung mutagenicity but not in fibrogenicity. Arch Toxicol. 2001;74:716-9.
22. Ustündağ A, Behm C, Föllmann W, Duydu Y, Degen GH. Protective effect of boric acid on lead- and cadmium-induced genotoxicity in V79 cells. Arch Toxicol. 2014;88:1281-9.
23. Venkov P, Topashka-Ancheva M, Georgieva M, Alexieva V, Karanov E. Genotoxic effect of substituted phenoxyacetic acids. Arch Toxicol. 2000;74:560-6.
24. Waisberg M, Joseph P, Hale B, Beyersmann D. Molecular and cellular mechanisms of cadmium carcinogenesis. Toxicology. 2003;192:95-117.
25. Westphal GA, Bünger J, Schulz TG, Müller MM, Hallier E. Mutagenicity of N-nitrosodiethylamine in the Ames test with S. typhimurium TA1535 is due to volatile metabolites and is not dependent on cytochrome P4502E1 induction. Arch Toxicol. 2000;74:638-41.
26. Zellmer S, Schmidt-Heck W, Godoy P, Weng H, Meyer C, Lehmann T, et al. Transcription factors ETF, E2F, and SP-1 are involved in cytokine-independent proliferation of murine hepatocytes. Hepatology. 2010;52:2127-36.
27. Zhang C, Lai Y, Jin G, Glatt H, Wei Q, Liu Y. Human CYP2E1-dependent mutagenicity of mono- and dichlorobiphenyls in Chinese hamster (V79)-derived cells. Chemosphere. 2015;144:1908-15.