Development of a K562 cell-based assay for screening anticancer agents  

以K562细胞为靶点的抗肿瘤药物筛选模型的建立(英文)

作　　者：钱静[1] 周彩红[1] 钱蓁 南发俊[1] 叶其壮 

机构地区：[1]中国科学院上海生命科学院上海药物研究所国家新药筛选中心,上海中国201203

出　　处：《Acta Pharmacologica Sinica》2001年第9期821-826,共6页中国药理学报（英文版）

基　　金：Project supported by the Ministry of Science & Technology of China (No 96-901-02-02);the National Natural Science Foundation of China (No 39725032);Shanghai Municipal Science Committee(No 99JC14049)

摘　　要：AIM: To develop a leukemia cell line K562-based assay for high-throughput screening. METHODS: The screening was carried out on 96-well plates with monitoring cell proliferation by a combined 3- [ 4, 5-dimethylthiazol-2-yl ] -5- [3-carboxymethoxyphenyl ] -2- [ 4-sulfophenyl]-2H-tetrazolium (MTS)/phenazine metho-sulfate (PMS) method. Conditions for evaluating effects on the proliferation of K562 cells by individual compounds on the 96-well plates were optimized. RESULTS: A set of 800 small organic compounds was screened for anticancer activity by this cell-based assay, with consumption of each compound at 500 ng. Eleven compounds were identified with > 80 % inhibitory activity at 5 mg/L, among which 9 compounds were confirmed by subsequent testing at multiple concentrations . The most potent compound showed an IC50 at 170 nmol/L, and there were total of 7 compounds showed IC50 less than 10 μmol/L. CONCLUSION: The high-throughput method using K562 cell line is fast, economical, effective, and practical in identifying inhibitors as potential therapeutic agents for cancer.目的:建立以白血病细胞系K562为靶点的高通量抗肿瘤药物筛选模型。方法:在96孔细胞培养板上,运用四唑氮化合物(MTS)和电子耦联剂(PMS)连用的方法,对K562细胞增殖情况进行检测。对在化合物影响下K562细胞增殖变化的检测条件进行了优化。结果:采用这一细胞水平的高通量抗肿瘤药物筛选模型,完成了800个小分子有机化合物的筛选,每个化合物的用量是500ng。11个化合物在浓度为5mg/L时可抑制细胞增殖达80％以上,其中9个通过多浓度复筛得到了确认。抑制活性最强的化合物的IC_(50)为170nmol/L,共有7个化合物显示IC_(50)低于10μmol/L。结论:采用K562细胞系进行高通量筛选是快速、经济、有效、实用的发现新型抗肿瘤药物的方法。

关 键 词：antitumor drug screening assays K562 cells cell culture drug screening antineoplastic agents 

分 类 号：R96[医药卫生—药理学]