Comparison of alamar blue and MTT assays for high through-put screening
Comparison of alamar blue and MTT assays for high through-put screening
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DOI：
10.1016/j.tiv.2004.03.012
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发表时间：
2004-10-01
影响因子：
3.2
通讯作者：
Bullock, P
中科院分区：
文献类型：
--
作者：
Hamid, R;Rotshteyn, Y;Bullock, P
比较了高通量形式的阿利西蓝和3-[4，5-二甲基噻唑-2-基]-2，5-二苯基溴化四唑（MTT）细胞活力测定的性能。在人肝癌细胞系HepG 2中使用这两种测定法在10 μ M下筛选了总共117种因其广泛的治疗领域而选择的药物。除了特非那定和阿司咪唑，这在两个试验中表现一致，阿替尼蓝测定法比MTT测定法对大多数化合物的灵敏度略高。MTT法检测柔红霉素和三氟拉嗪的效果不太敏感。阿司咪唑、柔红霉素、椭圆藤碱、氟奋乃静、特非那定、硫利达嗪和三氟拉嗪等七种药物在单点筛选的阿替尼蓝测定中的活力百分比结果为55%或更低。在两项试验中对这些进行了重新检测，以重新确认细胞毒性和测定EC 50值。除柔红霉素外，两种试验的EC 50值相当。基于这些结果和试验质量的Z因子评估，两种试验均为在候选药物选择的早期阶段鉴定体外细胞毒性药物提供了有用的信息。然而，由于代谢酶的诱导剂和/或抑制剂可能导致假阳性或假阴性结果，因此需要对数据进行仔细解释，这些代谢酶负责细胞毒性终点的转化，正如我们使用双香豆素所证明的那样。(C)2004爱思唯尔有限公司保留所有权利。
The performance of alamar blue and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) cell viability assays in a high through-put format were compared. A total of 117 drugs chosen for their wide range of therapeutic areas were screened at 10 muM using both assays in human hepatoma cell line HepG2. Except for terfenadine and astemizole, which performed consistently in both assays, the alamar blue assay was slightly more sensitive than the MTT assay for most compounds. The MTT assay was less sensitive detecting an effect for daunorubicin and trifluoperazine. Seven drugs, astemizole, daunorubicin, ellipticine, fluphenazine, terfenadine, thioridazine and trifluoperazine, had percent viability results of 55% or less in the alamar blue assay at the single point screen. These were re-tested in both assays for reconfirmation of cytotoxicity and determination of the EC50 values. Except for daunorubicin, the EC50 values were comparable in both assays. Based on these results and the Z-factor assessment of assay quality, both assays provided useful information to identify in vitro cytotoxic drugs at early stages of drug candidate selection. However, careful interpretation of data is warranted due to the possibility of false positive or negative results caused by inducers and/or inhibitors of metabolic enzymes that are responsible for transformation of cell toxicity end points, as we demonstrated using dicumarol. (C) 2004 Elsevier Ltd. All rights reserved.