To investigate the effect of interference of high-mobility-group protein A2 (HMGA2) on the in-vitro proliferation and invasion of HL-60 cells. Methods: RNA interference was used to inhibit HMGA2 expression by HL-60 cells. Animal model was established by inoculating nude mice with HL-60 cells, which was stably transfected with HMGA2-specific shRNA expression vector, and cells transfected with blank vector were used as negative control and cells without transfection as blank control. At different time points after inoculation, the percentage of leukemic cells in peripheral blood (PB) and bone marrow (BM) was detected by Wright’s staining and spleen and liver index were calculated at day 40 after inoculation. Results: RT-PCR and Western blot showed that HMGA2 expression by HL-60 cells was markedly reduced. BM smear demonstrated that HL-60 cells transfected with or without vectors were successfully implanted into nude mice. At day 21, 28, 40 after inoculation, significantly lower percentage of leukemic cells in PB and BM was seen in mice inoculated with HMGA2-targeted HL-60 cells than that in mice from both negative and blank control (P<0.05). The increase of leukemic cells in PB and BM along with prolonged time after inoculation was markedly slowed in HMGA2-targeted mice as compared with the other two groups. The complications occurred of mice in the experimental group, such as moving slowing, the abdominal swelling and the subcutaneous nodules were significantly delayed. The liver and spleen index determined in HMGA2-targeted mice were respectively 66.76±5.56 and 20.57±0.75, both which were remarkably lower than that in mice from the other groups (P<0.05). Conclusion: Interference of HMGA2 expression could remarkably inhibited HL-60 cell proliferation and invasion in nude mice.