Date of Award

4-2008

Degree Name

Master of Science

Department

Chemistry

First Advisor

Dr. Jay Means

Second Advisor

Dr. Elke Schoffers

Third Advisor

Dr. Susan Stapleton

Fourth Advisor

Dr. Laura Lowe Furge

Access Setting

Masters Thesis-Open Access

Abstract

The p53 protein has been documented to be mutated in more than 50% of late stage bladder cancers and tobacco causes 50% of all bladder tumors. The 4- arninobiphenyl (ABP) carcinogen comprises 5 ng per cigarette in the smoke and is a known bladder carcinogen. The research goal in the present study was to compare the reactivity of rat liver S-9 fraction activated 4-arninobiphenyl with the 45-mer sense strand of the TP53 exon7 gene to the respective anti-sense strand. The specific and sensitive technique of LC-tandem mass spectrometry (LC-MS/MS) was used for quantitation. The authentic standard of N-(adenine-8yl-)-4-arninobiphenyl (A-ABP) adduct was prepared by Buchwald-Hartwig coupling, used for MS tuning and LC-MS/MS calibration. The LC-MS/MS (n = 4, p = 0.10) results were 6 ± 3 and 10 ± 6 fmol on the column or reaction results of 5 ± 2 and 9 ± 4 N-( deoxyadenosine-8-yl)-4-aminobiphenyl ( dA-ABP) adducts / 106 nucleotides in the oligo for the sense and anti-sense strands, respectively. There were 8 vs. 5 dA and 9 vs. 4 dG non-hydrogen-bonded purines available for the sense and anti-sense strands, respectively, in the computationally folded oligos. The decreased reactivity measured for the sense strand is corroborated by computationally derived ssDNA secondary structure results that suggest that the sense strand ratio of available reactive dA to dG loci is less when compared to the anti-sense strand.

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