The Study On Differentiation Of Acute Myeloid Leukemia HL-60 Cells Induced By Puerariae Radix Flavones

【Objective】Puerarin is one of the major components of Xue-Fu-Kang created by the Affiliated Hospital of Nanjing University of Chinese Medicine. Prior researches have shown that Puerariae radix flavones (PRF) with the concentration of 50～200μg/ml, which is the active components of puerariae radix, could inhibit the proliferation of 4 sub-types cell lines (HL-60, Kasumi-1, NB4 and U937 respectively) of acute myeloid leukemia (AML). Lower concentration of PRF (50μg/ml) might induce HL-60 cells to differentiation, while 200μg/ml PRF could induce HL-60 to apotosis. This study aim to explore the differentiation of HL-60 cell line treated with the lower concentration of PRF (less than 50μg/ml).【Methods】(1) Treated with the PRF concentration from 10-100μg/ml for 24,48 and 72 hours, the cells proliferation inhibitory rates were analyzed by MTT assay.(2) Treated with10,30,50μg/ml PRF for 48 hours, the morphology changes and the percentage of NBT positive cells were determined microscopically counting by Wright's staining.(3) Treated with 10,30,50μg/ml PRF for 48 hours, cell surface differentiation-related antigenes CD11b, CD14,CD15,CD13 andCD33, and cell-cycle distribution were evaluated by flow cytometry (FCM).【Results】10,30,50μg/ml PRF inhibited HL-60 cells proliferation in 48 hours respectively, the cells exhibited the morphologic changes of granulocyte-like under the light microscope. The elevated NBT-positive cells were observed in this model meanwhile. Cell cycle analyses showed that cells in G0-G1 phase increased significantly, while S phase decreased in a dose-dependent manner. The positive percentages of CD11 and CD15 increased while CD33 declined statistically, meanwile the CD14 and CD13 changed little detected by FCM.[Conclusion]Lower doses PRF might inhibit cells proliferation, induce the HL-60 cells to granulocyte-like differentiation, block cells cycle to progression. The detail mechanisms woulde be investigated further.