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

AIM: To develop a leukemia cell line K562-based assay for high-throughput s creening. METHODS : The screening was carried out on 96-well plates with mo nitoring cell proliferation by a combined 3-[4, 5-dimethylthiazol-2-yl]-5-[ 3-carboxymethoxyphenyl]-2-[4-sulfophenyl]-2H-tetrazolium (MTS)/phenazine me thosulfate (PMS) method. Conditions for evaluating effects on the prolifera tion of K562 cells by individual compounds on the 96-well plates were optim ized. RESULTS: A set of 800 small organic compounds was screened for antica ncer activity by this cell-based assay, with consumption of each compound a t 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 nmo l/L, and there were total of 7 compounds showed IC50 less than 10 mu mol/L. CONCLUSION: The high-throughput method using K562 cell line is fast, econo mical, effective, and practical in identifying inhibitors as potential ther apeutic agents for cancer.