As the newest identified member of the matrix metalloproteinase (MMP) family, the expression pattern and function of epilysin (MMP-28) are still not well understood. Although epilysin was found to play an evolutionarily conserved role in neural development, the expression and function of epilysin in malignant glioma are unknown. Therefore, the aim of the present study was to quantitatively evaluate the expression level of epilysin in glioblastoma (GBM) and its association with clinical outcome of patients. For this purpose, a total of 216 GBM specimens and 31 normal brain specimens were collected in the present study. Expression level of epilysin was determined by immunohistochemistry assay and immunoreactivity score system. MGMT promoter methylation and IDH1/2 mutation status in GBM were also evaluated. Results showed that the positive rate of epilysin staining in GBM was significantly elevated compared with that in normal brain. Positive epilysin staining was associated with low KPS score, unmethylated MGMT promoter and wild-type IDH. Kaplan-Meier analysis showed that patients with GBM of positive epilysin staining were more likely to have unfavorable overall survival. Multivariate analysis revealed that epilysin was an independent and significant prognostic marker of GBM. These results proved for the first time that epilysin expression was significantly elevated in GBM and can be potentially used to predict prognosis in patients with GBM.