title: Human formalin-fixed paraffin-embedded tissues: an untapped specimen for biomonitoring of carcinogen DNA adducts by mass spectrometry.
abstract: DNA adducts represent internal dosimeters to measure exposure to environmental and endogenous genotoxicants. Unfortunately, in molecular epidemiologic studies, measurements of DNA adducts often are precluded by the unavailability of fresh tissue. In contrast, formalin-fixed paraffin embedded (FFPE) tissues frequently are accessible for biomarker discovery. We report here that DNA adducts of aristolochic acids (AAs) can be measured in FFPE tissues at a level of sensitivity comparable to freshly frozen tissue. AAs are nephrotoxic and carcinogenic compounds found in Aristolochia herbaceous plants, many of which have been used worldwide for medicinal purposes. AAs are implicated in the etiology of aristolochic acid nephropathy and upper urinary tract carcinoma. 8-Methoxy-6-nitrophenanthro-[3,4-d]-1,3-dioxole-5-carboxylic acid (AA-I) is a component of Aristolochia herbs and a potent human urothelial carcinogen. AA-I reacts with DNA to form the aristolactam (AL-I)-DNA adduct 7-(deoxyadenosin-N(6)-yl) aristolactam I (dA-AL-I). We established a method to quantitatively retrieve dA-AL-I from FFPE tissue. Adducts were measured, using ultraperformance liquid chromatography/mass spectrometry, in liver and kidney tissues of mice exposed to AA-I, at doses ranging from 0.001 to 1 mg/kg body weight. dA-AL-I was then measured in 10-μm thick tissue-sections of FFPE kidney from patients with upper urinary tract cancers; the values were comparable to those observed in fresh frozen samples. The limit of quantification of dA-AL-I was 3 adducts per 10(9) DNA bases per 2.5 μg of DNA. The ability to retrospectively analyze FFPE tissues for DNA adducts may provide clues to the origin of human cancers for which an environmental cause is suspected.
authors.affiliation: Division of Environmental Health Sciences, Wadsworth Center, New York State Department of Health, Albany, New York 12201, United States.
authors.forename: Byeong Hwa
authors.initials: BH
authors.lastname: Yun
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Rosenquist
authors.forename: Jovan
authors.initials: J
authors.lastname: Nikolić
authors.forename: Dejan
authors.initials: D
authors.lastname: Dragičević
authors.forename: Karla
authors.initials: K
authors.lastname: Tomić
authors.forename: Bojan
authors.initials: B
authors.lastname: Jelaković
authors.forename: Kathleen G
authors.initials: KG
authors.lastname: Dickman
authors.forename: Arthur P
authors.initials: AP
authors.lastname: Grollman
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Turesky
mesh_terms: D000818:Animals; D034341:Aristolochic Acids / Q000737:chemistry; D002273:Carcinogens / Q000032:analysis*; D002853:Chromatography, Liquid; D018736:DNA Adducts / Q000032:analysis*; D005557:Formaldehyde / Q000737:chemistry*; D006801:Humans; D007680:Kidney Neoplasms / Q000473:pathology*; D008297:Male; D013058:Mass Spectrometry; D051379:Mice; D008810:Mice, Inbred C57BL; D015394:Molecular Structure; D016612:Paraffin Embedding; D016707:Tissue Fixation; D014516:Ureteral Neoplasms / Q000473:pathology*
chemical_list: D034341:Aristolochic Acids; D002273:Carcinogens; D018736:DNA Adducts; D005557:Formaldehyde
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