Objective To investigate the effects of co-treatment with puerariae radix flavones (PRF) and arsenic trioxide (ATO) on proliferation and apoptosis of Kasumi-1 cells and HL-60 cells of acute myeloid leukemia M2. Methods Kasumi-1 cells and HL-60 cells were treated with 100 μg/mL PRF and 1 μmol/L ATO for 48 h (RRF+ATO group). MTT assay was employed to measure the inhibition rate of cell proliferation, Annexin V-FITC/PI double staining was used to determine the early apoptosis rate by flow cytometry, the expression of Bcl-2 and Survivin mRNA was detected by Real-Time PCR, and the expression of Caspase-3, Caspase-8 and Caspase-9 protein was detected by Western blotting. Cells only treated with 100 μg/mL PRF were served as PRF group, and blank control group was also established. Results The inhibition rate of cell proliferation, early apoptosis rate and expression of Caspase-3 and Caspase-9 protein of Kasumi-1 cells in RRF+ATO group were significantly higher than those in PRF group (P<0.05), and there was an obvious trend of down-regulation of expression of Bcl-2 mRNA in Kasumi-1 cells. There was no significant difference in each parameter of HL-60 cells between RRF+ATO group and PRF group (P>0.05). Conclusion Co-treatment with PRF and ATO can effectively inhibit the proliferation and induce early apoptosis of Kasumi-1 cells, while that has no significant effect on HL-60 cells.