Type 2 interleukin-1 receptor (IL1R2) is a member of the IL1 family. It has been reported that IL1R2 is expressed and plays important roles in several cancers. However, its potential role in human osteosarcoma has not been reported. In the present study, a recombinant lentivirus harboring short hairpin RNA against IL1R2 was constructed and then transfected human osteosarcoma U-2 OS cells. The proliferation of infected cells was measured by 3-(4, 5- dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide (MTT) assay and plate colony formation assay. Results showed that efficiency of lentivirus infection was more than 80 %. The knockdown efficiency of IL1R2 mRNA level was up to 95 % compared with control group. After treatment with lentiviruses containing shIL1R2 (Lv-shIL1R2), the proliferation and colony formation of U-2 OS cells were obviously reduced (P < 0.001). Cell cycle assay was used to investigate the mechanism of the inhibition of proliferation of U-2 OS cells. We found that after Lv-shIL1R2 treatment, the percentage of cells at G0/G1 phase decreased significantly, whereas cells at S and G2/M phases increased markedly (P < 0.05). In conclusion, shIL1R2 indeed inhibited the proliferation of U-2 OS cells which might be associated with the blockage of S and G2/M phases. The results suggested that IL1R2 could have oncogenic potential and shIL1R2 might represent a new and effective therapeutic treatment for osteosarcoma patients.