Acute promyelocytic leukemia (APL) is a severe type of acute leukemia and the prognosis of patients was poor. Indirubin is the active constituent of the traditional Chinese medicine qingdai and an indoline anti-tumor drug. N’-Acetylindirubin is a novel indirubin derivative with better curative effect and less side effect. In this study, the effects of N’-Acetylindirubin on proliferation, apoptosis and cell cycle of acute myeloid leukemia cell line HL-60 was examined. The results demonstrated that N’-Acetylindirubin significantly induced apoptotic cell death in a dose and time-dependent manner and arrested cell cycle in G2/M in HL-60 cells. N’-Acetylindirubin also suppressed cyclin D1. This study suggests that N’-Acetylindirubin may serves as a potential chemopreventive agent for acute promyelocytic leukemia.