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Gewählte Publikation:

Publikationstyp: Zeitschriftenaufsatz
Dokumenttyp: Originalarbeit

Jahr: 1993

AutorInnen: Baird, WM; Lau, HH; Schmerold, I; Coffing, SL; Brozich, SL; Lee, H; Harvey, RG

Titel: Analysis of polycyclic aromatic hydrocarbon-DNA adducts by postlabelling with the weak beta-emitters 35S-phosphorothioate and 33P-phosphate, immobilized boronate chromatography and high-performance liquid chromatography.

Quelle: IARC Sci Publ. 1993; 152(124):217-226

Autor/innen der Vetmeduni Vienna:

Schmerold Ivo

Beteiligte Vetmed-Organisationseinheiten
Institut für Pharmakologie und Toxikologie

The wide range of carcinogenic activities of optical isomers of bay-region diol-epoxides of polycyclic aromatic hydrocarbons (PAHs) demands analytical techniques capable of distinguishing both the diastereoisomer and enantiomers of the PAH diol-epoxide (PAH-DE) responsible for DNA adduct formation. Our laboratory recently developed postlabelling procedures using the radioisotope 35S for HPLC analysis of benzo[a]pyrene (BaP)-DNA adducts formed in BaP-treated hamster cell cultures. To allow identification and complete separation of anti-PAH-DE-DNA adducts from syn-PAH-DE-DNA adducts, an immobilized boronate chromatography procedure was developed using a high-capacity boronate column. Analysis of 7,12-dimethyl-benz[a]anthracene (DMBA)-DNA adducts formed in treated hamster embryo cells by this boronate chromatography procedure and 35S-postlabelling demonstrated that both anti- and syn-DMBA-DE were bound to DNA. These analytical techniques also provided evidence that the potent carcinogen and environmental PAH dibenzo[a,l]pyrene was metabolized in cells to a DNA-binding intermediate with structural characteristics of an anti-PAH-DE. Conditions were also developed for using 33P-labelled ATP in the postlabelling assay. The use of [33P] instead of [35S]ATP offers the advantages of ease of labelling and greater sensitivity while still using a weak beta-emitter. The development of PAH-DNA adduct analysis techniques with improved sensitivity allows more detailed studies of how PAHs interact with DNA and how this leads to cancer induction.

Keywords Pubmed: 9,10-Dimethyl-1,2-benzanthracene/analysis
Cells, Cultured
Chromatography, High Pressure Liquid/methods
DNA/drug effects
DNA Adducts*
DNA Damage
Phosphorus Radioisotopes
Polycyclic Compounds/analysis*
Polycyclic Compounds/toxicity
Sulfur Radioisotopes

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