Background: The overall prognosis of brainstem gliomas is very poor, and the current treatment cannot significantly prolong the overall survival of these patients; therefore, studying the molecular biological mechanisms of the occurrence and development of brainstem gliomas has important significance for their treatment. The Wnt/beta-catenin signaling pathway is closely associated with the occurrence and development of tumors, but its relationship with brainstem gliomas remains unclear. Methods: This study used Western blot and immunohistochemistry methods to detect the expressions of Wnt/beta-catenin signaling pathway-related components such as Wnt-1, Wnt-2, beta-catenin and C-myc in six cases of normal brain tissues and 24 cases of brainstem gliomas and analyzed the relationship between their expressions and clinicopathological characteristics. Results: Wnt-1 had no obvious expression in normal brain tissues and did not show any significant difference between high-and low-grade brainstem gliomas; the expressions of Wnt-2, beta-catenin and C-myc in high-grade brainstem gliomas were significantly higher than that in low-grade brainstem gliomas and normal brain tissues and were positively correlated with the expression of Ki-67. Moreover, the expressions of Wnt-2 and C-myc were significantly associated with the prognosis of brainstem glioma patients; additionally, there was a trend toward increased beta-catenin expression with shorter survival, but there was no statistical difference. Conclusions: Wnt/beta-catenin signaling pathway might be abnormally activated and plays an important role in the occurrence and development of brainstem gliomas. Wnt-2, beta-catenin and C-myc may be potential targets for brainstem glioma treatment.