Determination of point mutational spectra of benzo[a]pyrene-diol epoxide in human cells
- Univ. of Pittsburgh, PA (United States)
- Massachusetts Institute of Technology, Cambridge, MA (United States)
The primary goal of our research consists of developing means sufficiently sensitive to allow assessment of human exposure to environmental carcinogens. We describe here a new approach for analyzing point mutational spectra and a test for its validity and precision using cultured human cells exposed to high doses of environmental carcinogens. The approach in its present form includes (a) treatment of independent large cultures of human cells with a carcinogen, (b) selection of mutant cells en masse by 6-thioguanine resistance, (c) amplification of sequences of interest directly from 6TG[sup R] cells using high-fidelity polymerase chain reaction, and (d) separation of mutant sequences from nonmutant sequences using denaturing gradient gel electrophoresis. We report use of this protocol to observe induced mutational spectra in exon 3 of the hprt gene in cultured human cells by benzo[a]pyrene-diol epoxide (BPDE), an active form of the widely distributed environmental carcinogen benzo[a]pyrene. BPDE induced predominantly G to T transversions within this target sequence. The variation of the frequency of the mutations among independent cultures is consistent with the interpretation that each of them corresponds to a hotspot. 17 refs., 4 figs.
- DOE Contract Number:
- FG02-86ER60448
- OSTI ID:
- 6848508
- Journal Information:
- Environmental Health Perspectives; (United States), Journal Name: Environmental Health Perspectives; (United States) Vol. 98; ISSN EVHPAZ; ISSN 0091-6765
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
560300* -- Chemicals Metabolism & Toxicology
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
AROMATICS
BENZOPYRENE
BIOLOGICAL HOT SPOTS
CARCINOGENESIS
CONDENSED AROMATICS
DNA SEQUENCING
ELECTROPHORESIS
HYDROCARBONS
MUTATIONS
ORGANIC COMPOUNDS
PATHOGENESIS
STRUCTURAL CHEMICAL ANALYSIS