Reduction of misleading ("false") positive results in mammalian cell genotoxicity assays. III: Sensitivity of human cell types to known genotoxic agents
Reduction of misleading ("false") positive results in mammalian cell genotoxicity assays. III: Sensitivity of human cell types to known genotoxic agents
复制标题
DOI：
10.1016/j.mrgentox.2014.03.001
复制
发表时间：
2014-06-01
影响因子：
1.9
通讯作者：
Pfuhler, Stefan
中科院分区：
文献类型：
--
作者：
Fowler, Paul;Smith, Robert;Pfuhler, Stefan
我们之前已经证明，当使用啮齿动物来源的细胞系和某些毒性指标（如相对细胞计数或复制指数）时，体外微核试验（MNvit）的“假”或“误导性”阳性结果的发生率似乎更高。这些研究表明，使用具有功能性p53的人类细胞类型和包括细胞增殖功能的毒性测量可以显著减少误导性阳性结果的检测。降低“假阳性率”不应该以丧失检测灵敏度为代价。因此，我们研究了MNvit检测试剂盒对已知遗传毒性试剂的灵敏度，使用了之前显示不太容易出现误导性阳性的三种细胞类型，即人淋巴细胞（HuLy）、TK6和HepG2细胞。这17种化学物质的特征得到了很好的表征，并且来自已知在体外哺乳动物细胞测定中产生阳性结果的化学物质列表。这些数据证明了采用TK6和HuLy细胞的测定的高灵敏度，使得17种化学物质中的15种被正确鉴定。总之，优先使用TK6和HuLy细胞而不是长期建立的啮齿动物细胞系以提高特异性不会损害MNvit检测已知遗传毒性剂的灵敏度。(C)2014爱思唯尔有限公司版权所有。
We have demonstrated previously that the seemingly high rate of "false" or "misleading" positive results from in vitro micronucleus assays (MNvit) was greater when rodent derived cell lines and certain toxicity measures, such as relative cell count or replication index, were used. These studies suggested that the use of a human cell type with functional p53 and a toxicity measure that included a function of cell proliferation could dramatically reduce the detection of misleading positive results.A reduced "false positive rate" should not be at the expense of a loss of sensitivity of the assay. Therefore, we have investigated the sensitivity of the MNvit assay to known genotoxic agents using three cell types shown previously to be less prone to misleading positives, namely human lymphocytes (HuLy), TK6 and HepG2 cells. The 17 chemicals are well characterised and are from a list of chemicals known to produce positive results in in vitro mammalian cell assays.These data demonstrated a high sensitivity of the assay in which TK6 and HuLy cells were employed, such that 15 out of the 17 chemicals were correctly identified. By contrast, the use of HepG2 cells resulted in far fewer than expected positive responses.In conclusion, using TK6 and HuLy cells in preference to long established rodent cell lines in order to improve specificity does not compromise the sensitivity of the MNvit to detect known genotoxic agents. (C) 2014 Elsevier B.V. All rights reserved.