Glioblastoma (GBM) remains the most aggressive brain tumor, characterized by rapid proliferation and resistance to temozolomide (TMZ). Emerging evidence suggests that the Wnt signaling through stabilization of β-catenin and expression of the non-canonical ligand Wnt5a, has been implicated in the maintenance of GBM. Cannabidiol (CBD) has emerged as a potential adjuvant that may modulate these pathways to overcome chemoresistance. Therefor we determined human U87-MG cells treatment with Different concentration of CBD and TMZ determined in 24, 48, and 72hours. Cell viability, combination index, IC₅₀and IC₅ were assed with MTT assay. Protein levels of β-catenin, Wnt5a, and AXIN1 were evaluated using western blotting and immunofluorescence assay. Treatment with CBD in combination TMZ significantly reduced U87-MG cell viability in a time and dose dependent manner. The combination index analysis indicated a synergistic interaction between CBD and TMZ, particularly at 48 and72 hours. IC₅₀values decreased significantly enhanced co administration compared to mono therapy, suggesting enhanced chemosensitivity. Western blotting and IF results revealed that down regulation of β-catenin and Wnt5a by increasing AXIN1 expression, supporting modulation of canonical and non-canonical Wnt signaling pathways. These findings demonstrate that CBD potentiates the cytotoxic effects of TMZ in glioblastoma cells by interfering with Wnt/β-catenin signaling pathway may represent a promising adjuvant strategy to improve therapeutic outcomes in GBM.