Matrix metalloproteinases (MMP) have been identified as biomarkers for several diseases, including cancer. MMP-26 is constitutively expressed in some cancer cells of epithelial origin. Despite this, there is a lack of studies regarding the expression of MMP-26 on prostatic carcinoma. Here, we investigate the expression of the MMP-26 peptide in benign and malign prostatic tissues. For this, 150 specimens, including atrophy (N = 25), prostatic intraepithelial neoplasia (PIN) (N = 25), benign prostatic hyperplasia (BPH) (N = 50), and prostatic adenocarcinoma (PA) (N = 50), were immunohistochemically (IHC) examined for the expression of MMP-26. MMP-26 expression was positive in 70 (46.7%) out of the 150 samples, being more prevalent in the PA group (46/50 cases,92%), followed by PIN (22/25 cases, 88%). The BPH group showed only 2/50 (4%) positive cases, and the atrophy group showed no reactivity. ROC curve analysis showed that MMP-26 immunoexpression had a higher area under the curve between PA vs atrophy+PIN+BPH (AUC=0.94; 95% CI 0.9-0.98), PA+PIN vs atrophy+BPH (AUC=0.97; 95% CI 0.94-0.99) and PA vs atrophy+BPH (AUC=0.97; 95% CI 0.95-1.00) groups. In addition, the expression and intensity of the MMP-26 reaction showed a significant association with total PSA values (P=0.001). Our results showed that MMP-26 immunoexpression was useful to differentiate a group of benign and malignant samples in prostate tumors. This characteristic could assist in the predictive assessment and, consequently, in the development of new strategies for the diagnosis, prognosis, and treatment of prostate cancer.