Postlabeling methods for carcinogen-DNA adduct analysis

Journal Article · Tue Oct 01 04:00:00 EDT 1985 · Environ. Health Perspect.; (United States)

DOI:https://doi.org/10.1289/ehp.856257· OSTI ID:5319907

Randerath, K; Randerath, E; Agrawal, H P; Gupta, C; Schurdak, M E; Vijayaraj, M

Radioactive carcinogens have provided most of our present knowledge about the chemistry of interactions between carcinogens and biological systems. To extend the scope of carcinogen-DNA binding studies, an alternative method, which does not require radioactive test chemicals, has been developed. In this approach, radioactivity (/sup 32/P) is being incorporated into DNA constituents by polynucleotide kinase-catalyzed (/sup 32/P)phosphate transfer from (..gamma..-/sup 32/P)ATP after exposure of the DNA in vitro or in vivo to a nonradioactive, covalently binding chemical, and evidence for the alteration of DNA nucleotides is provided by the appearance of extra spots on autoradiograms of thin-layer chromatograms of digests of the chemically modified DNA. Quantitation of adduct levels is accomplished by scintillation counting. The sensitivity of the technique depends on the experimental conditions for /sup 32/P-labeling and on the chemical structure of the adducts. A total of approximately 80 compounds has been studied thus far. Binding to DNA of rodent tissues was readily detected by the /sup 32/P-postlabeling assay for all known carcinogens among these compounds, and adducts were detected in DNA from human placenta of smokers.

• Research Organization: Baylor College of Medicine, Houston, TX
• OSTI ID: 5319907
• Journal Information: Environ. Health Perspect.; (United States), Journal Name: Environ. Health Perspect.; (United States) Vol. 62; ISSN EVHPA
• Country of Publication: United States
• Language: English

Related Subjects

550601* -- Medicine-- Unsealed Radionuclides in Diagnostics
62 RADIOLOGY AND NUCLEAR MEDICINE
ADDUCTS
ANIMALS
ATP
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARCINOGENS
CHROMATOGRAPHY
COUNTING TECHNIQUES
DAYS LIVING RADIOISOTOPES
DNA ADDUCTS
ENZYMES
ISOTOPES
LABELLING
LIGHT NUCLEI
MAMMALS
NUCLEI
NUCLEOTIDES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
PHOSPHOTRANSFERASES
RADIOISOTOPES
RODENTS
SCINTILLATION COUNTING
SEPARATION PROCESSES
THIN-LAYER CHROMATOGRAPHY
TRANSFERASES
VERTEBRATES