Glioblastoma (GBM) is the most aggressive and treatment-resistant primary brain tumor in adults. Despite current multimodal therapies, including surgery, radiation, and temozolomide chemotherapy, patient outcomes remain poor. Enhancing tumor radiosensitivity through biocompatible nanomaterials could provide a promising integrative strategy for improving therapeutic effectiveness. This study aims to evaluate the potential of bovine serum albumin-coated copper oxide nanoparticles (BSA@CuO-NPs) to enhance radiosensitivity in U87-MG cells under clinically relevant X-ray irradiation. In brief, BSA@CuO-NPs were synthesized via carbodiimide crosslinking and characterized by DLS, SEM, and zeta potential analysis. U87-MG cells were treated with BSA@CuO-NPs alone or in combination with X-ray irradiation (2 Gy). Cytotoxicity was assessed using the MTT assay, while radiosensitization was evaluated through clonogenic survival analysis. Apoptosis induction and DNA damage were analyzed via Annexin V staining and γ-H2AX immunofluorescence, respectively. The results revealed that BSA@CuO-NPs showed good colloidal stability and biocompatibility compared with uncoated CuO-NPs. When combined with irradiation, BSA@CuO-NPs significantly decreased clonogenic survival (p < 0.05) and increased apoptotic cell death compared to irradiation alone. Immunofluorescence demonstrated increased γ-H2AX focus formation, indicating higher DNA double-strand breaks in the combination group. In conclusion, BSA@CuO-NPs enhance the effects of ionizing radiation by increasing DNA damage and apoptosis in U87-MG cells, indicating their potential as combined radiosensitizers. These results support further research into albumin-coated metal oxide nanoparticles as adjuncts to standard radiotherapy for the management of GBM. One challenge in this context is the effective delivery of nanoparticles to GBM. However, the stability of BSA@CuO-NPs in physiological solutions could help overcome this obstacle.