以K562细胞为靶点的抗肿瘤药物筛选模型的建立

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，运用四唑氯化合物（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IC50为170nmol/L，共有7个化合物显示IC50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。

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

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 methosulfate (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 micromol/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.