Dehydroepiandrosterone sulphate (DHEAS) has a protective role against cancer; however, the mechanism by which DHEAS has this effect remains poorly understood. The purpose of this study was to investigate the potential effects of DHEAS on HL-60 (human promyelocytic leukemia cell). The percentage of cells undergoing apoptosis was analyzed by flow cytometry using Annexin V-fluorescein isothiocyanate staining. DHEAS internalization into HL-60 cells was visualized by transmission electron microscopy. DHEAS could be used to facilitate uptake to specific cancer cells for cancer therapy and diagnosis. Our results showed that the uptake of DHEAS by HL-60 cells were also much higher than that of the control cells. In this study, the mechanism of DHEAS actions against leukemia involves the inhibition of cell proliferation indicating that DHEAS can be useful in the treatment of leukemia.