Pinhal Danillo, Gontijo Alisson Marques de Miranda Cabral, Reyes Victor Alexis Valenzuela, Salvadori Daisy Maria Favero
Núcleo de Avaliação Toxicogenética e Cancerígena (TOXICAN), Departamento de Patologia, Faculdade de Medicina, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil.
Environ Mol Mutagen. 2006 Mar;47(2):117-26. doi: 10.1002/em.20174.
Buccal mucosa (BM) cells have been used in human biomonitoring studies for detecting DNA adducts and chromosomal damage in an epithelial cell population. In the present study, we have investigated if human BM cells are suitable for use in the single-cell gel electrophoresis (SCGE)/Comet assay as an approach for estimating the exposure of epithelial cells to DNA-damaging agents. Our results indicate that only a few cells from BM cell samples yield comets that can be analyzed by current methods, and that the yield of cells with comets is independent of the percentage of viable BM cells in the sample. Data generated after enzymatic enrichment of viable cells and immunomagnetic separation of epithelial cells suggest that most of the BM cells that do form comets are probably leukocytes. Moreover, by reevaluating specific cells after running the Comet assay, we found that viable epithelial BM cells give rise to atypical comets that are not included in the analysis. Comparing DNA migration patterns between small groups of smokers and nonsmokers indicated that long-term smoking had no effect on the subpopulation of cells that yield typical comets. Our results indicate that the SCGE assay, as it is commonly performed, may not be useful for genotoxicity monitoring in human epithelial BM cells.
颊黏膜（BM）细胞已被用于人类生物监测研究，以检测上皮细胞群体中的DNA加合物和染色体损伤。在本研究中，我们调查了人类BM细胞是否适合用于单细胞凝胶电泳（SCGE）/彗星试验，作为评估上皮细胞暴露于DNA损伤剂的一种方法。我们的结果表明，BM细胞样本中只有少数细胞产生的彗星可以用当前方法进行分析，并且有彗星的细胞产量与样本中活BM细胞的百分比无关。对活细胞进行酶富集和上皮细胞免疫磁珠分离后产生的数据表明，大多数形成彗星的BM细胞可能是白细胞。此外，通过在彗星试验后重新评估特定细胞，我们发现活的上皮BM细胞会产生不包括在分析中的非典型彗星。比较一小群吸烟者和非吸烟者之间的DNA迁移模式表明，长期吸烟对产生典型彗星的细胞亚群没有影响。我们的结果表明，通常进行的SCGE试验可能对人类上皮BM细胞的遗传毒性监测没有用处。
