Cytoskeletal-associated protein 2 (CKAP2), which is also known as tumor-associated microtubule-associated protein, has been reported to be dysregulated in various types of human cancer. However, the role of CKAP2 in glioma has not been fully elucidated. The present study evaluated the expression pattern of CKAP2 using the Chinese Glioma Genome Atlas microarray database, which included 301 patients, and validated the findings using The Cancer Genome Atlas RNA sequencing database. Kaplan-Meier survival analysis, and univariate and multivariate Cox analyses, were used to estimate survival distributions. Furthermore, the biological implication of aberrant CKAP2 expression in high-grade glioma (HGG) was investigated using Gene Ontology analysis, gene set enrichment analysis, gene set variation analysis and STRING. The results indicated that patients with HGG exhibited significantly higher CKAP2 expression levels compared with patients with low-grade glioma in both databases. Higher expression levels of CKAP2 were significantly associated with shorter overall survival and progression-free survival of patients with HGG. Furthermore, CKAP2 was also positively correlated with known malignant factors, including high Ki67 expression and phosphatase and tensin homolog mutations. The univariate and multivariate Cox regression analyses demonstrated that CKAP2 may be a novel independent prognostic biomarker for patients with HGG. Functional assays also indicated that CKAP2 was closely associated with the cell cycle, mitosis and cell proliferation. These results suggested that CKAP2 may be associated with tumor growth and could serve as an independent prognostic factor, particularly in patients with HGG.