Speit Günter, Schütz Petra, Högel Josef, Schmid Oliver
Universität Ulm, Institut für Humangenetik, D 89069 Ulm, Germany.
Mutagenesis. 2007 Nov;22(6):387-94. doi: 10.1093/mutage/gem031. Epub 2007 Sep 13.
Formaldehyde (FA) is known to be genotoxic and mutagenic in proliferating mammalian cells in vitro. The present study was performed to further characterize its genotoxic potential in the V79 Chinese hamster cell line. The induction of DNA strand breaks and DNA-protein cross-links (DPXs) was measured by the comet assay in relationship to the induction of sister chromatid exchanges (SCEs) and micronuclei (MN). Induction of DNA strand breaks was found neither with the standard protocol of the alkaline comet assay nor with modifications using extended electrophoresis times or proteinase K. The concentration-effect relationship for the genotoxic effects was characterized by fitting different curves to the data. A two-phase regression model fitted best in comparison with a linear or a quadratic model and indicated practical thresholds for the induction of SCE and MN. For the induction of DPX as measured by the comet assay, neither a linear concentration-response relationship nor any of the tested models fitted well to the data. Three repeated treatments with genotoxic concentrations of FA with a 3-h interval led to enhanced levels of DPX and MN while the same treatments with a 24-h interval did not enhance FA genotoxicity but suggested adaptive protection against the DNA-damaging action of FA.
已知甲醛(FA)在体外增殖的哺乳动物细胞中具有遗传毒性和致突变性。本研究旨在进一步表征其在V79中国仓鼠细胞系中的遗传毒性潜力。通过彗星试验测量DNA链断裂和DNA-蛋白质交联(DPX)的诱导情况,并与姐妹染色单体交换(SCE)和微核(MN)的诱导情况相关联。无论是碱性彗星试验的标准方案,还是使用延长电泳时间或蛋白酶K的改进方法,均未发现DNA链断裂的诱导。通过将不同曲线拟合到数据中来表征遗传毒性效应的浓度-效应关系。与线性或二次模型相比,两相回归模型拟合效果最佳,并表明了SCE和MN诱导的实际阈值。对于通过彗星试验测量的DPX诱导,线性浓度-反应关系或任何测试模型均不能很好地拟合数据。用遗传毒性浓度的FA进行三次重复处理,间隔3小时,导致DPX和MN水平升高,而间隔24小时进行相同处理并未增强FA的遗传毒性,但表明对FA的DNA损伤作用具有适应性保护。