Department of Radiation Genetics, Graduate School of Medicine, Kyoto University, Yoshidakonoe, Sakyo-ku, Kyoto 606-8501, Japan.
DNA Repair (Amst). 2010 Dec 10;9(12):1292-8. doi: 10.1016/j.dnarep.2010.09.017. Epub 2010 Oct 27.
Chemicals used industrially and commercially are required by law to be assessed for their genotoxic potential. However, all currently used assays have major limitations and despite intense effort, there is no universal agreement on which tests should be employed, or how to interpret results. We have developed a new assay system using the chicken DT40 B cell line that offers a number of significant advantages over current methodologies. Our assay could provide enhanced sensitivity using genetically defined and phenotypically characterized mutants defective in DNA repair pathways. Furthermore, analysis of the mutants, using DNA repair proficient wild-type cells as a negative control, minimizes false negative outcomes. Assessing the different responses of a panel of mutants representative of all repair pathways, mechanistic detail of genotoxicity can be determined. This unique feature, as well as reducing the false positive rate, strengthens positive identifications and is useful when extrapolating results to the human context. Our panel of mutants is likely to be useful in screening large compound libraries for an emerging class of chemotherapeutic drugs, which includes inhibitors of DNA repair enzymes such as PARP and DNA polymerases.
工业和商业中使用的化学物质，根据法律规定，需要对其遗传毒性进行评估。然而，目前所有使用的检测方法都存在重大局限性，尽管进行了大量努力，但对于应该使用哪些检测方法或如何解释结果，尚无普遍共识。我们使用鸡 DT40 B 细胞系开发了一种新的检测系统，该系统与当前方法相比具有许多显著优势。我们的检测方法可以使用在 DNA 修复途径中具有遗传定义和表型特征的突变体来提高灵敏度。此外，使用 DNA 修复功能正常的野生型细胞作为阴性对照来分析突变体，可以最大限度地减少假阴性结果。通过分析代表所有修复途径的一组突变体的不同反应，可以确定遗传毒性的机制细节。这种独特的功能，以及降低假阳性率，增强了阳性鉴定，并且在将结果推断到人类环境时非常有用。我们的突变体面板可能有助于筛选新兴的一类化疗药物（包括 DNA 修复酶如 PARP 和 DNA 聚合酶的抑制剂）的大型化合物文库。
