Novel biomarkers of genetic damage in humans: use of fluorescence in situ hybridization to detect aneuploidy and micronuclei in exfoliated cells.
Moore-LE; Titenko-Holland-N; Quintana-PJ; Smith-MT
J Toxicol Environ Health 1993 Oct; 40(2-3):349-357
New biomarkers of genetic damage in humans were developed and validated for use in exposed populations. Urine was collected from healthy nonsmoking volunteers, aged 23 to 42 years. Buccal cells were collected from the inside of the mouth. Nasal cells were also obtained. Fluorescence in-situ hybridization (FISH) was used with centromeric and chromosome specific DNA probes to detect aneuploidy and micronucleus formation in exfoliated human epithelial cells. The new fluorescent technique gave results that were very similar to those obtained with the standard Feulgen fast green method. The spontaneous levels of micronuclei in healthy volunteers were: buccal, 0.13%; nasal, 0.21%; and urothelial, 0.07%. These were based on approximately 1500 cells per data point. The values were lower than that found in cultured lymphocytes, 0.4 to 0.8%. About 50% of the exfoliated cell micronuclei contained whole chromosomes. FISH was also used to detect aneuploidy in exfoliated buccal and bladder cells. The estimated frequency of aneuploidy in exfoliated cells was similar to that found in human lymphocytes analyzed by FISH with the same probe for chromosome 9. The authors conclude that FISH can be used to characterize micronuclei present in exfoliated epithelial cells and to measure spontaneous levels of aneuploidy. The use of FISH was being expanded to include additional gene and chromosome specific probes. Preliminary trials using these assays in epidemiologic studies of exposed populations were in progress.
NIOSH-Publication; NIOSH-Grant; Training; Genotoxic-effects; Urinalysis; DNA-damage; Chromosome-damage; Mutagenesis; Biological-monitoring
Biomedical & Environ Hlth Scis University of California 322 Warren Hall Berkeley, Calif 94720
Journal of Toxicology and Environmental Health
University of California Berkeley, Berkeley, California