This study is to investigate the mechanism of human promyelocytic leukemia HL-60 cells proliferation induced by alteronol in vitro.  Human promyelocytic leukemia HL-60 cells cultured in vitro were treated with different concentrations of alteronol.  Inhibition rate was detected by SRB assay.  Cellular morphological changes were observed by Hoechst and AO/EB (acridine orange/ethidium bromide dye) staining.  The apoptosis rate was determined by Annexin V-FITC/PI assay.  Cell cycle distribution was determined by flow cytometry.  Western blotting analysis was carried out to determine the cell cycle related proteins.  The proliferation of HL-60 cells treated with alteronol was inhibited in a concentration-dependent manner.  Based on cell viability assay, observation on cell morphology and apoptosis rate, it confirmed that alteronol played an obvious role in proliferation inhibition of human promyelocytic leukemia HL-60 cells, but it did not induce apoptosis in human promyelocytic leukemia HL-60 cells in different concentrations groups.  Alteronol could effectively inhibit the proliferation of human promyelocytic leukemia HL-60 cells inducing cell cycle arrest at G₁ phase, as well as, alteration expression of cell cycle proteins level of CyclinD1 and pRb.