The Further Development of a Mammalian DNA Alkaline Unwinding Bioassay With Potential Application to Hazard Identification for Contaminants from Environmental Samples

Recently, we have detailed a DNA alkaline unwinding assay (DA UA) that can be used to rapidly measure chemically induced strand breaks in mammalian cells (Daniel et al., 1985). In this paper we present further development of this assay, including: (1) studies on the relationship between DNA adducts...

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Bibliographic Details
Published in:Toxicology and industrial health 1989-10, Vol.5 (5), p.647-665
Main Authors: Daniel, F. Bernard, Chang, Lina W., Schenck, Kathleen M., Deangelo, Anthony B., Skelly, Michael F.
Format: Article
Language:English
Subjects:
560300 - Chemicals Metabolism & Toxicology
ADDUCTS
ANIMALS
AROMATICS
BENZOPYRENE
BIOASSAY
BIOLOGICAL EFFECTS
BIOSYNTHESIS
BODY
CARCINOGEN SCREENING
Carcinogenicity Tests - methods
CELL CULTURES
Cells, Cultured
CONDENSED AROMATICS
DIGESTIVE SYSTEM
DNA
DNA ADDUCTS
DNA Damage
Environmental Pollutants - toxicity
FLUORENE
GENETIC EFFECTS
GLANDS
Humans
HYDROCARBONS
Hydrogen-Ion Concentration
LIVER
Liver - cytology
Male
MAMMALS
MAN
METABOLIC ACTIVATION
MICE
MUTAGEN SCREENING
Mutagenicity Tests - methods
NUCLEIC ACIDS
ORGANIC COMPOUNDS
ORGANS
PRIMATES
RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT
RODENTS
SCREENING
Spleen - cytology
STRAND BREAKS
SYNTHESIS
VERTEBRATES
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Summary:Recently, we have detailed a DNA alkaline unwinding assay (DA UA) that can be used to rapidly measure chemically induced strand breaks in mammalian cells (Daniel et al., 1985). In this paper we present further development of this assay, including: (1) studies on the relationship between DNA adducts and DNA strand breaks; (2) evaluation of the role of cytotoxicity in DNA strand breaks; and (3) application of the DA UA to cell preparations from the liver of mice dosed with methylating agents. The level of DNA adducts produced in human CCRF-CEM cells by treatment with benzo(a)pyrene diol-epoxide (BPDE), N-acetoxy-2-acetyl amino fluorene (AAAF), and various methylating agents was linear with concentration over several orders of magnitude. Likewise, the level of strand breaks increased with the concentration over the same dose range. The strand breaks/adduct ratio ranged from 0.05 for the methyl adducts to 0.001 for the BPDE adducts. Using these values and the inherent sensitivity of the DA UA (circa 100 to 1000 breaks/ cell), (Daniel et al., 1985), the ability of the assay to detect DNA damage induced by various classes of chemical carcinogens can be calculated. The DA UA appears to be useful for assessing the relative potency of various environmental genotoxic effects on mammalian cells. In addition, it can be conducted on cells isolated from target organs of whole animals.
ISSN:0748-2337
1477-0393
DOI:10.1177/074823378900500506