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Effect of Staining Procedures on the Results of Micronucleus Assays with Exfoliated Oral Mucosa Cells
Ist Teil von
Cancer epidemiology, biomarkers & prevention, 2006-10, Vol.15 (10), p.1835-1840
Ort / Verlag
Philadelphia, PA: American Association for Cancer Research
Erscheinungsjahr
2006
Quelle
MEDLINE
Beschreibungen/Notizen
Micronuclei in exfoliated epithelial cells are widely used as biomarkers of cancer risk in humans. To elucidate the effect
of different staining procedures on the outcome of such investigation, we conducted a study in which the micronuclei frequencies
in oral mucosa cells of heavy smokers ( n = 20) and nonsmokers ( n = 10) were evaluated with nonspecific (Giemsa, May-Grünwald-Giemsa) and DNA-specific (4′,6-diamidino-2-phenylindole, Feulgen,
acridine orange) stains, whereas with Giemsa-based stains, the frequencies of micronuclei in smokers were significantly (4-
to 5-fold) higher in the smokers group, no significant increase was observed with any of the DNA-specific stains. Furthermore,
the evaluation of cells of the two study groups with Feulgen stain showed that oral mucosa cells from smokers had significantly
increased levels of nuclear anomalies other than micronuclei. These anomalies are consequences of cell injury found in epithelial
cells and are paralleled by formation of keratin bodies in the cytoplasm that resemble micronuclei. Correlation analyses showed
that micronuclei frequencies scored in Giemsa-stained slides correlated significantly with karyorrhexis, karyolysis, condensed
chromatin, and binucleates, whereas no such correlations were found with DNA-specific stains. These findings indicate that
nuclear anomalies (and possibly keratin bodies) may be misinterpreted as micronuclei with nonspecific DNA stains and lead
to false-positive results in studies with cells of epithelial origin. Furthermore, our results show that exposure of oral
mucosa cells to genotoxic carcinogens contained in tobacco smoke does not lead to induction of micronuclei in these cells.
(Cancer Epidemiol Biomarkers Prev 2006;15(10):1835–40)