FEASIBILITY OF A HIGH-FLUX ANTICANCER DRUG SCREEN USING A DIVERSE PANEL OF CULTURED HUMAN TUMOR-CELL LINES
FEASIBILITY OF A HIGH-FLUX ANTICANCER DRUG SCREEN USING A DIVERSE PANEL OF CULTURED HUMAN TUMOR-CELL LINES
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DOI：
10.1093/jnci/83.11.757
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发表时间：
1991-06-05
期刊：
影响因子：
--
通讯作者：
BOYD, M
中科院分区：
文献类型：
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作者：
MONKS, A;SCUDIERO, D;BOYD, M
我们在这里描述了一个中试规模的开发和实施，在体外，抗癌药物筛选利用一组60人肿瘤细胞系组织成代表白血病，黑色素瘤，肺癌，结肠癌，肾癌，卵巢癌和中枢神经系统的子面板。 这种疾病导向筛选的最终目标是促进发现具有潜在细胞系特异性和/或亚组特异性抗肿瘤活性的新化合物。 在当前筛选方案中，将各细胞系接种到微量滴定板上，然后预孵育24-28小时。 随后，以5个10倍稀释液加入测试试剂，并将培养物再孵育48小时。 对于每种测试试剂，生成剂量-反应曲线。 通过原位固定细胞，然后用蛋白结合染料磺酰罗丹明B（SR B）染色，进行细胞活力或细胞生长的终点测定。 SRB与细胞大分子的碱性氨基酸结合;溶解的染色剂通过荧光光度法测定，以确定处理和未处理细胞中的相对细胞生长或活力。 在试点筛选研究之后，每周400种化合物的筛选率一直保持不变。
We describe here the development and implementation of a pilot-scale, in vitro, anticancer drug screen utilizing a panel of 60 human tumor cell lines organized into subpanels representing leukemia, melanoma, and cancers of the lung, colon, kidney, ovary, and central nervous system. The ultimate goal of this disease-oriented screen is to facilitate the discovery of new compounds with potential cell line-specific and/or subpanel-specific antitumor activity. In the current screening protocol, each cell line is inoculated onto microtiter plates, then preincubated for 24-28 hours. Subsequently, test agents are added in five 10-fold dilutions and the culture is incubated for an additional 48 hours. For each test agent, a dose-response profile is generated. End-point determinations of the cell viability or cell growth are performed by in situ fixation of cells, followed by staining with a protein-binding dye, sulforhodamine B (SRB). The SRB binds to the basic amino acids of cellular macromolecules; the solubilized stain is measured spectrophotometrically to determine relative cell growth or viability in treated and untreated cells. Following the pilot screening studies, a screening rate of 400 compounds per week has been consistently achieved.