The expression of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of matrix metalloproteinase-3 (TIMP-3) in giant-cell tumor of bone (GCT), and the correlation of their expression with the clinicopathologic features and prognosis were investigated. A total of 70 GCT patients treated in our hospital from September, 2013 to September, 2015, were selected, and the tumor and para-carcinoma tissues were obtained by surgery. The expression levels of MMP-2 and TIMP-3 in GCT and para-carcinoma tissues were detected via semi-quantitative polymerase chain reaction (PCR) and western blot analysis, as well as immunohistochemical staining. Moreover, the clinicopathological data of the GCT patients were collected to study the correlation of MMP-2 and TIMP-3 with clinicopathological features and prognosis of GCT. The results of semi-quantitative PCR and western blot analysis revealed that the expression level of MMP-2 in tissues of the 70 GCT patients was significantly higher than that in para-carcinoma tissues, and the difference was statistically significant (P<0.01), while the expression level of TIMP-3 was obviously lower than that in para-carcinoma tissues (P<0.01). The results of immunohistochemical staining revealed that the positive expression rate of MMP-2 was 57.6% in GCT tissues and 18.9% in para-carcinoma tissues, while that of TIMP-3 was 63.2% in GCT tissues and 13.8% in para-carcinoma tissues, and the differences were statistically significant (P<0.01). The expression levels of MMP-2 and TIMP-3 were correlated with the diameter of tumor, clinical staging, lymph node metastasis and relapse of GCT (P<0.01), but were not correlated with the age and sex of GCT patients (P>0.05). There was a negative correlation between MMP-2 and TIMP-3 expression levels (r=−0.258, P<0.05). The expression levels of MMP-2 and TIMP-3 are closely related to the clinicopathological features and prognosis of patients, which can be used as one of the clinical examination indexes of GCT and also provide new insights for the clinical treatment of GCT.