title: Semiquantitation of polycyclic aromatic hydrocarbon-DNA adducts in human esophagus by immunohistochemistry and the automated cellular imaging system.
abstract: It has been suggested that ingestion of polycyclic aromatic hydrocarbons (PAHs) may contribute to the high incidence and mortality of esophageal cancer in Linxian, China. To explore this relationship a semiquantitative immunohistochemical staining method was developed for localization of PAH-DNA adducts. Nuclear color intensity (bright field average pink intensity per nucleus for >1000 cells) was measured using the ChromaVision Automated Cellular Imaging System (ACIS). Paraffin-embedded sections of cultured human keratinocytes exposed to increasing concentrations of 7beta,8alpha-dihydroxy-9alpha,10alpha-epoxy-7,8,9,10-tetrahydro-benzo[a]pyrene (BPDE) were incubated with BPDE-DNA antiserum and served as an internal positive control (standard curve). Values for nuclear staining intensity correlated directly with BPDE exposure concentration (r(2) = 0.99) and were reproducible. DNA adduct levels determined by BPDE-DNA chemiluminescence immunoassay in DNA from BPDE-exposed keratinocytes, correlated with BPDE exposure concentrations (r(2) = 0.99), showing that nuclear staining intensity determined by ACIS correlated directly with BPDE-DNA adduct levels determined by chemiluminescence immunoassay. The ACIS methodology was applied to 5 human samples from Linxian, and significantly positive nuclear PAH-DNA adduct staining was observed in this group when compared with esophageal tissue from 4 laboratory-housed monkey controls and 6 samples obtained at autopsy from smokers and nonsmokers in the United States. Nuclear PAH-DNA staining was absent from Linxian samples when serial sections were incubated with normal rabbit serum (negative control) and was significantly reduced on incubation with BPDE-DNA antiserum absorbed previously with the immunogen BPDE-DNA. These results appear to support the hypothesis that high PAH exposure levels may be etiologically associated with the development of esophageal cancer in Linxian.
authors.affiliation: Laboratory of Cellular Carcinogenesis and Tumor Promotion, Division of Cancer Treatment and Diagnosis, National Cancer Institute, NIH, Bethesda, Maryland 20892, USA.
authors.forename: Hilde E
authors.initials: HE
authors.lastname: van Gijssel
authors.forename: Rao L
authors.initials: RL
authors.lastname: Divi
authors.forename: Ofelia A
authors.initials: OA
authors.lastname: Olivero
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Roth
authors.forename: Guo-Qing
authors.initials: GQ
authors.lastname: Wang
authors.forename: Sanford M
authors.initials: SM
authors.lastname: Dawsey
authors.forename: Paul S
authors.initials: PS
authors.lastname: Albert
authors.forename: You-Lin
authors.initials: YL
authors.lastname: Qiao
authors.forename: Philip R
authors.initials: PR
authors.lastname: Taylor
authors.forename: Zhi-Wei
authors.initials: ZW
authors.lastname: Dong
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Schrager
authors.forename: David E
authors.initials: DE
authors.lastname: Kleiner
authors.forename: Miriam C
authors.initials: MC
authors.lastname: Poirier
mesh_terms: D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide / Q000494:pharmacology*; D000818:Animals; D001331:Automation; D002294:Carcinoma, Squamous Cell / Q000453:epidemiology / Q000473:pathology* / Q000601:surgery; D002681:China / Q000453:epidemiology; D003470:Culture Media; D046508:Culture Techniques; D018736:DNA Adducts / Q000032:analysis / Q000378:metabolism* / Q000494:pharmacology*; D003952:Diagnostic Imaging / Q000379:methods*; D004938:Esophageal Neoplasms / Q000453:epidemiology / Q000473:pathology* / Q000601:surgery; D016629:Esophagectomy; D006801:Humans; D007150:Immunohistochemistry; D015603:Keratinocytes / Q000187:drug effects* / Q000473:pathology*; D011084:Polycyclic Aromatic Hydrocarbons / Q000032:analysis / Q000378:metabolism*; D011817:Rabbits; D012016:Reference Values; D015203:Reproducibility of Results; D012494:Sampling Studies; D012680:Sensitivity and Specificity
chemical_list: D003470:Culture Media; D018736:DNA Adducts; D011084:Polycyclic Aromatic Hydrocarbons; C043931:benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide-DNA; C095911:polycyclic aromatic hydrocarbons-DNA adduct; D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide