title: Altered gene expression in mouse livers after dichloroacetic acid exposure.
abstract: Dichloroacetic acid (DCA) is a major by-product of water disinfection by chlorination. Several studies have demonstrated that DCA exhibits hepatocarcinogenic effects in rodents when administered in drinking water. This chemical does not appear to be highly mutagenic, and the mechanism(s) involved in DCA induction of cancer are not clear. The present work was aimed at identifying changes in gene expression which may indicate critical alterations/pathways involved in this chemical's carcinogenic activities. We used cDNA microarray methods for analyses of gene expression in livers of mice treated with the tumorigenic dose of 2 g/l DCA in drinking water for 4 weeks. Total RNA samples obtained from livers of the control and DCA-treated mice were evaluated for gene expression patterns with Clontech Atlas Mouse 1.2 cDNA and Atlas mouse stress/toxicology arrays, and the data analyzed with AtlasImage 2.01 and one-way ANOVA in JMP4 software. From replicate experiments, we identified 24 genes with altered expression, of which 15 were confirmed by Northern blot analysis. Of the 15 genes, 14 revealed expression suppressed two- to five-fold; they included the following: MHR 23A, cytochrome P450 (CYP) 2C29, CYP 3A11, serum paraoxonase/arylesterase 1 (PON 1), liver carboxylesterase, alpha-1 antitrypsin, ER p72, glutathione S-transferase (GST) Pi 1, angiogenin, vitronectin precursor, cathepsin D (CTSD), plasminogen precursor (contains angiostatin), prothrombin precursor and integrin alpha 3 precursor (ITGA 3). An additional gene, CYP 2A4/5, had a two-fold elevation in expression. Further, in ancillary Northern analyses of total RNA isolated from DCA-induced hepatocellular carcinomas (from earlier reported studies of mice treated with 3.5 g/l DCA for 93 weeks), many of the same genes (11 of 15) noted above showed a similar alteration in expression. In summary, we have identified specific genes involved in the functional categories of cell growth, tissue remodeling, apoptosis, cancer progression and xenobiotic metabolism that have altered levels of expression following exposures to DCA. These findings serve to highlight new pathways in which to further probe DCA effects that may be critical to its tumorigenic activity.
authors.affiliation: Environmental Carcinogenesis Division, National Health and Environmental Effects Research Laboratory, Mail Drop 68, US Environmental Protection Agency, Research Triangle Park, NC 27711, USA. thai.sheau-fung@epa.gov
authors.forename: Sheau-Fung
authors.initials: SF
authors.lastname: Thai
authors.forename: James W
authors.initials: JW
authors.lastname: Allen
authors.forename: Anthony B
authors.initials: AB
authors.lastname: DeAngelo
authors.forename: Michael H
authors.initials: MH
authors.lastname: George
authors.forename: James C
authors.initials: JC
authors.lastname: Fuscoe
mesh_terms: D000818:Animals; D003999:Dichloroacetic Acid / Q000009:adverse effects*; D005786:Gene Expression Regulation; D008099:Liver / Q000378:metabolism*; D008114:Liver Neoplasms, Experimental; D008297:Male; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000032:analysis; D015262:Xenobiotics / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D015262:Xenobiotics; D003999:Dichloroacetic Acid