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The aim of this study was to evaluate the role of matrix metalloproteinases (MMPs), their tissue inhibitors [tissue inhibitors of metalloproteinases (TIMPs)] and activators [membrane-type MMPs (MT1-MMPs)], vascular endothelial growth factor (VEGF) and endostatin on clinicopathological variables and prognosis in patients with rectal cancer.Paired samples of tumour tissue and normal tissue were obtained from patients with rectal cancer who underwent curative surgery (n = 34). Gelatin zymography for MMP-2 and MMP-9, an activity assay for MT1-MMP and enzyme-linked immunoassays for TIMP-2, VEGF and endostatin were performed using extracts from the paired tissue samples.Active MMP-9 showed statistically significant relationships with metastatic disease and perineural invasion (P = 0.002 and P = 0.042). A significant relationship was observed between the levels of tumoral pro-MMP-2 and pro-MMP-9 and the presence of lymph node metastasis (P = 0.012 and P = 0.021, respectively). Tumoral TIMP-2 levels showed a significant relationship with tumour recurrence (P = 0.011). A significant relationship was also observed between tumour VEGF levels and the presence of perineural invasion (P = 0.044), and VEGF levels were correlated with the size of the tumour (P = 0.009, r = 0.454).These results might contribute to further investigation of a possible prognostic significance in rectal cancer.