/sup 32/P-postlabeling analysis of aromatic DNA adducts in fish from polluted areas

Journal Article · Tue Dec 15 04:00:00 EST 1987 · Cancer Res.; (United States)

OSTI ID:5601516

Dunn, B P; Black, J J; Maccubbin, A

Brown bullheads (Ictalurus nebulosus) were sampled from sites in the Buffalo and Detroit Rivers where fish are exposed to high levels of sediment bound polycyclic aromatic hydrocarbons, and suffer from an elevated frequency of liver cancer. DNA was isolated from the livers of these wild fish and from control specimens which were raised in clean aquariums. DNA was enzymatically digested to normal and adducted nucleotides, and hydrophobic/bulky adducts were enriched in the digests either by preparative reverse-phase high-pressure liquid chromatography, or selective nuclease P1 dephosphorylation of normal nucleotides. Aromatic DNA-carcinogen adducts were then quantitated using /sup 32/P-postlabeling analysis. Using both adduct enrichment procedures, chromatograms derived from DNA of fish from polluted areas showed a diffuse diagonal radioactive zone not present in DNA from aquarium raised fish. The diagonal zone appeared to consist at least in part of multiple overlapping discrete adduct spots which could be partially separated by gradient high-pressure liquid chromatography prior to /sup 32/P-postlabeling analysis, and most of which were more strongly retained on a reverse-phase column than the major benzo(a)pyrene-DNA adduct. The behavior of the adducts in the diagonal radioactive zone and of their unlabeled precursors is consistent with their identification as nucleotide adducts of a variety of bulky hydrophobic aromatic environmental compounds. Total pollution-related adduct levels as analyzed by HPLC adduct enrichment and /sup 32/P-postlabeling were 70.1 +/- 29 (SD) nmol/mol normal nucleotide in fish from the Buffalo River, and 52 and 56 nmol/mol for two specimens from the Detroit River.

• Research Organization: British Columbia Cancer Research Centre, Vancouver
• OSTI ID: 5601516
• Journal Information: Cancer Res.; (United States), Journal Name: Cancer Res.; (United States) Vol. 47; ISSN CNREA
• Country of Publication: United States
• Language: English

Related Subjects

550501 -- Metabolism-- Tracer Techniques
560300* -- Chemicals Metabolism & Toxicology
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ADDUCTS
ANIMALS
AQUATIC ORGANISMS
AROMATICS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CARCINOGENESIS
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
DETROIT RIVER
DIGESTIVE SYSTEM
DISEASES
DNA
DNA ADDUCTS
FEDERAL REGION V
FISHES
GLANDS
HYDROCARBONS
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
LIQUID COLUMN CHROMATOGRAPHY
LIVER
MICHIGAN
NEOPLASMS
NORTH AMERICA
NUCLEI
NUCLEIC ACIDS
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANS
PATHOGENESIS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
POLLUTION
POLYCYCLIC AROMATIC HYDROCARBONS
RADIOISOTOPES
RIVERS
SEPARATION PROCESSES
STREAMS
SURFACE WATERS
TRACER TECHNIQUES
USA
VERTEBRATES
WATER POLLUTION