We investigated the expression of CXC chemokine receptor 4 (CXCR4), vascular endothelial growth factor (VEGF) and matrix metalloproteinase (MMP)-9 to elucidate whether these markers could predict lymph node metastasis in human breast cancer. Higher rates of CXCR4 (61%), VEGF (68%), and MMP-9 (63%) expression were found in breast cancer tissues than in normal and atypical hyperplasia tissues. The expression of these markers was significantly associated with primary tumor progression, histological grade, and lymph node status. We found there were significant correlations between the expressions of any two of the three markers (P<0.001). Furthermore, our studies indicated that concomitant expression of CXCR4/VEGF (P=0.007), CXCR4/MMP-9 (P<0.001) or VEGF/MMP-9 (P=0.003) had stronger correlation with lymph node metastasis than did each alone and that combined expression of all three makers strongly correlated with lymph node metastasis (P<0.001). Thus, simultaneously examining the expression of CXCR4, VEGF, and MMP-9 in cancer tissues of breast cancer will provide valuable prognostic diagnosis of lymph node metastasis.