Chemokine (C-X-C motif) receptor 4 (CXCR4) has been found to closely correlate with the incidence, development, treatment and prognosis of breast cancer. The aim of the present study was to investigate the effects of CXCR4 on bone metastasis in breast cancer and to explore the mechanisms of this process. CXCR4 small interfering RNA was transfected into the breast cancer cell line, MDA-MB-231BA-rfp, and the cell proliferation and invasion abilities of the cells were measured using cell counting kit-8 cell proliferation and Transwell assays. A mouse model of breast cancer with bone metastasis was prepared and the bone metastasis was confirmed using micro-positron emission tomography. The associated proteins were detected by western blot analysis and the results showed that CXCR4 RNAi inhibited the cell proliferation and invasion ability of the MDA-MB-231BA-rfp cells. In addition, CXCR4 RNAi inhibited the duration and extent of bone metastasis in the MDA-MB-231BA-rfp cells in the mouse model, while the inhibition of CXCR4RNAi blocked the phosphatidylinositide 3-kinase (PI3K)/protein kinase B (AKT)/matrix metalloproteinase (MMP)-9 pathway. In conclusion, the present study demonstrated that CXCR4 RNAi inhibits bone metastasis and the cell proliferation and invasion abilities of breast cancer cells. Furthermore, the CXCR4/PI3K/AKT/MMP-9 pathway may be important in the bone metastasis of breast cancer.