COMPARATIVE SYNCHRONOUS FLUORESCENCE SPECTROPHOTOMETRY AND P-32 POSTLABELING ANALYSIS OF PAH-DNA ADDUCTS IN HUMAN LUNG AND THE RELATIONSHIPTO TP53 MUTATIONS

Polycyclic aromatic hydrocarbon (PAH)-DNA adducts were studied in huma n lung from 39 lung cancer patients by synchronous fluorescence spectr ophotometric (SFS) and P-32-postlabelling assays. Regression analysis of the samples failed to detect any correlation between benzo[a]pyrene -diolepoxide (BPDE)-DNA adducts detected by SFS and the BPDE co-migrat ing spot detected by P-32-postlabelling. We have also analyzed the rel ationship between adduct levels and TP53 mutations. By postlabelling d iagonal radioactive zone (DRZ) adducts were detected in 37 of 39 (95%) lung tissues from lung cancer patients and the adduct level ranged fr om 6.81 to 108.50 adducts/10(8) nucleotide, Thirty-three of 39 (85%) h ad detectable levels of BPDE-DNA adducts (>1 adduct/10(9) nucleotide). Current heavy smokers (>20 cigarettes/day) have significantly higher DRZ adduct levels compared to individuals smoking less than 20 cigaret tes/day. By SFS combined with immunoaffinity column (IAC), 11 of 39 (2 8%) samples had detectable adduct levels, and 6 of 11 (55%) were detec table by SFS following purification of benzo[a]pyrene (BP)-tetrols by high pressure liquid chromatography (HPLC), Six of 33 (18%) samples we re positive for BPDE-DNA adducts by both postlabelling and HPLC/SFS, N o correlation was observed between the SFS and P-32-postlabelling assa ys for the detection of BPDE-DNA adducts, However, there was a good co rrelation between adduct levels detected by IAC/SFS and HPLC/SFS, We f ound a weak association between total PAH-DNA adduct levels in lung ti ssue and TP53 mutations.