Correlation between mutation, gene expression and DNA adduct formation in human lymphoblast cells dosed with benzo[a]pyrene diol epoxide

TOXI 57

Caroline Ceailles, carolineceailles@gmail.com1, Wen Luo, wendyluo@alum.mit.edu2, Wenhong Fan, wfan@fhcrc.org2, Lue P Zhao, lzhao@fhcrc.org2, Helmut Zarbl, zarbl@eohsi.rutgers.edu3, and Paul Vouros, p.vouros@neu.edu1. (1) Department of Chemistry and Chemical Biology, Northeastern University, 360 Huntington Ave, Boston, MA 02115, (2) Fred Hutchinson Cancer Research Center, Seattle, WA 98109, (3) Environmental and Occupational Health Sciences Institute, UMDNJ-Robert Wood Johnson Medical School, Piscataway, NJ 08854
There is substantial evidence showing that benzo[a]pyrene (B[a]P), a major polyaromatic hydrocarbon found in cigarette smoke, contributes to carcinogenesis via the formation of mutagenic compounds generated through metabolic activation. Nonetheless, a complete understanding of its chemical and biological effects is still lacking. This study combines the analysis of DNA adducts in human TK6 lymphoblastoid cells after exposure to the mutagenic metabolite benzo[a]pyrene-7,8-diol-9,10-epoxide (B[a]PDE) with a comprehensive assessment of the cellular responses as a function of exposure. An LC-MS/MS assay was developed for the detection and quantification of B[a]PDE-N2-deoxyguanosine (B[a]PDE-N2-dG), the principal DNA adduct of B[a]P. The assay was used to measure the adduct levels in human TK6 lymphoblastoid cells as a function of dose and time after exposure to B[a]PDE. The levels of B[a]PDE-N2-dG adducts were then correlated to cell toxicity, induced mutation at the TK (thymidine kinase) and HPRT loci, and gene expression profiling through microarray analysis.

 

Poster Session and Awards
6:00 PM-10:00 PM, Tuesday, August 21, 2007 BCEC -- 204 A/B, Poster

Division of Chemical Toxicology

The 234th ACS National Meeting, Boston, MA, August 19-23, 2007