The aim of this work was to investigate the expression and prognostic significance of microtubule-associated serine/threonine kinase 1 (MAST1) in glioma tissues. MAST1 mRNA expression in tumor tissues and prognosis of patients with gliomas were analyzed with the GEPIA 2 database. mRNA seq_325 and mRNA seq_693 transcriptome data and clinical data were obtained from the Chinese Glioma Genome Atlas (CGGA) database, differential expression of MAST1 mRNA in glioma tissues with different clinical features was analyzed. Impact of MAST1 expression on prognosis of patients with glioma was evaluated using Kaplan-Meier survival curves and multivariate Cox regression analysis. Potential signaling pathways involved in MAST1 were explored by Gene Set Enrichment Analysis (GSEA) software. MAST1 mRNA expression was found to be low in glioma tissues. MAST1 mRNA expression was correlated with patient age, WHO grade, IDH mutation, and 1p/19q co-deletion status. Low MAST1 expression was found associated with poor prognosis in glioma patients. Further enrichment analysis revealed involvement of MAST1 expression in multiple biological processes. We conclude that MAST1 is a potential indicator for prognostic evaluation in glioma patients.