title: Differentiation of DNA reactive and non-reactive genotoxic mechanisms using gene expression profile analysis.
abstract: Genotoxic stress triggers a variety of biological responses including the transcriptional activation of genes regulating DNA repair, cell survival and cell death. Here, we investigated whether gene expression profiles can differentiate between DNA reactive and DNA non-reactive mechanisms of genotoxicity. We analyzed gene expression profiles and micronucleus levels in L5178Y cells treated with cisplatin and sodium chloride. The assessment of cisplatin genotoxicity (up to six-fold increase in the number of micronuclei) and gene expression profile (increased expression of genotoxic stress-associated genes) was in agreement with cisplatin mode of action as a DNA adduct-forming agent. The gene expression profile analysis of cisplatin-treated cells identified a number of genes with robust up regulation of mRNA expression including genes associated with DNA damage (i.e. members of GADD45 family), early response (i.e. cFOS), and heat shock protein (i.e. HSP40 homologue). The gene expression changes correlated well with DNA damage as measured by DNA-protein crosslinks and platinum-DNA binding. To differentiate the genotoxic stress-associated expression profile of cisplatin from a general toxic stress, we have compared the gene expression profile of cisplatin-treated cells to cells treated with sodium chloride, which causes osmotic shock and cell lysis. Although the sodium chloride treatment caused a two-fold induction of micronuclei, the gene expression profile at equitoxic concentrations was remarkably distinct from the profile observed with cisplatin. The profile of sodium chloride featured a complete lack of expression changes in genes associated with DNA damage and repair. In summary, the gene expression profiles clearly distinguished between DNA reactive and non-reactive genotoxic mechanisms of cisplatin and sodium chloride. Our results suggest the potential utility of gene expression profile analysis for elucidating mechanism of action of genotoxic agents.
authors.affiliation: Pfizer Global Research and Development Eastern Point Rd, Groton, CT 06340, USA.
authors.forename: Donna A
authors.initials: DA
authors.lastname: Dickinson
authors.forename: Gregory R
authors.initials: GR
authors.lastname: Warnes
authors.forename: George
authors.initials: G
authors.lastname: Quievryn
authors.forename: Joseph
authors.initials: J
authors.lastname: Messer
authors.forename: Anatoly
authors.initials: A
authors.lastname: Zhitkovich
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Rubitski
authors.forename: Jiri
authors.initials: J
authors.lastname: Aubrecht
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D020869:Gene Expression Profiling; D008223:Lymphoma / Q000235:genetics; D051379:Mice; D009153:Mutagens / Q000633:toxicity*
chemical_list: D009153:Mutagens