Toxicity of folic acid analogs in cultured human cells: a microtiter assay for the analysis of drug competition.
Toxicity of folic acid analogs in cultured human cells: a microtiter assay for the analysis of drug competition.
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叶酸类似物在培养的人体细胞中的毒性：用于分析药物竞争的微量滴定测定。
DOI：
10.1073/pnas.84.14.4860
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发表时间：
1987
影响因子：
11.1
通讯作者：
R. Schimke
中科院分区：
文献类型：
--
作者：
D. Roos;R. Schimke
我们已经使用了微量滴定法来研究各种叶酸类似物在一系列培养的人类细胞系中的毒性，这些细胞系对氨甲喋呤（一种二氢叶酸还原酶抑制剂）表现出不同程度的抗性。尽管二氢叶酸还原酶基因扩增，这些细胞仍保持其对亲脂性抗叶酸剂BW301U的敏感性。由于研究中的细胞系生长非常缓慢，并且在非条件培养基中的平板接种效率很差，因此开发了一种依赖于细胞增殖而不是集落形成作为毒性量度的测定法。这种方法很容易推广，以提供一个快速和廉价的药物竞争测定。二维研究表明，甲氨蝶呤和BW301U的毒性，竞争和亚叶酸救援模式的差异，表明这两种药物作用于不同的目标。进一步应用微量滴定分析的多药物相互作用的分析进行了讨论。
We have used a microtiter assay to study the toxicity of various folate analogs in a series of cultured human cell lines that exhibit different degrees of resistance to methotrexate, an inhibitor of dihydrofolate reductase. These cells retain their sensitivity to the lipophilic antifolate BW301U despite the amplification of dihydrofolate reductase genes. Because the cell lines under investigation grow very slowly and have poor plating efficiencies in unconditioned medium, an assay was developed that relies on cell proliferation rather than colony formation as a measure of toxicity. This approach is easily generalized to provide a rapid and inexpensive assay of drug competition. Two-dimensional studies indicate that methotrexate and BW301U show differences in patterns of toxicity, competition, and rescue by folinic acid, suggesting that the two drugs act on different targets. Further applications of the microtiter assay to the analysis of multidrug interactions are discussed.