Rapid detection of DNA-damaging agents using repair-deficient CHO cells

C. A. Hoy, E. P. Salazar, L. H. Thompson

Research output: Contribution to journalArticle

71 Citations (Scopus)

Abstract

A screening method is introduced to detect and claissify DNA-damaging agents using DNA repair-deficient strains of Chinese hamster ovary cells. Differential cytotoxicity (relative growth) of the mutant cells compared to the wild-type cells was interpreted as a measure of lethal, potentially repairable damage to DNA. The assay consists of exposing the wild-type cells and three mutant strains to the test compound in a 24-well tray and using staining intensity to estimate growth after 72 h. The battery of mutants consists of two UV-sensitive strains (UV4 and UV5) that are deficient in different aspects of nucleotide excision repair, and strain EM9, which is defective in DNA-strand-break rejoining. The assay was highly reproducible, and the magnitude of the differential cytotoxicity response compared favorably with the amount of differential killing measured by colony-formation survival curves for several chemicals. 15 direct-acting and 7 metabolism-dependent agents that were expected to produce bulky, covalent DNA adducts were tested in the assay, and all produced a differential cytotoxicity response in at leastwo of the test UV5 showed a response to all of the test compounds whereas EM9 showed a response to 7 of the test compounds. Thus, the pattern of mutant responses presumably reflects the types of DNA damage produced by a compound. Although this aspect is still under development, these results indicate the potential of a larger battery of mutants to classify a wide spectrum of chemicals according to the lesions they produce. 13 non-DNA damaging agents were also tested and none produced a differential cytotoxicity response, suggesting that this endpoint is specific for DNA damage. We conclude that this assay may be a cost-effective alternative or adjunct to the existing short-term tests.

Original languageEnglish (US)
Pages (from-to)321-332
Number of pages12
JournalMutation Research/Environmental Mutagenesis and Related Subjects
Volume130
Issue number5
DOIs
StatePublished - 1984
Externally publishedYes

Fingerprint

CHO Cells
Repair
Cytotoxicity
DNA Damage
Assays
DNA
DNA Repair-Deficiency Disorders
DNA Breaks
DNA Adducts
Growth
Cricetulus
DNA Repair
Ovary
Staining and Labeling
Metabolism
Costs and Cost Analysis
Screening
Nucleotides
Cells
Costs

ASJC Scopus subject areas

  • Genetics
  • Toxicology
  • Medicine(all)

Cite this

Rapid detection of DNA-damaging agents using repair-deficient CHO cells. / Hoy, C. A.; Salazar, E. P.; Thompson, L. H.

In: Mutation Research/Environmental Mutagenesis and Related Subjects, Vol. 130, No. 5, 1984, p. 321-332.

Research output: Contribution to journalArticle

Hoy, C. A. ; Salazar, E. P. ; Thompson, L. H. / Rapid detection of DNA-damaging agents using repair-deficient CHO cells. In: Mutation Research/Environmental Mutagenesis and Related Subjects. 1984 ; Vol. 130, No. 5. pp. 321-332.
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