Glioblastoma is the most aggressive malignancy of the human brain, accounting for 40% of all primary malignant brain tumors. However, there is no effective treatment for this disease. This study was designed to develop anti-vascular endothelial growth factor (VEGF) as a novel adjuvant therapy for glioblastoma. A VEGF shRNA vector was constructed to silence VEGF expression in U251 glioma cells and these cells were treated with various concentrations of liposomal paclitaxel, 6 Gy radiation or liposomal paclitaxel plus radiation. The data demonstrated that the VEGF shRNA vector significantly knocked down VEGF expression, which synergistically sensitized U251 glioma cells to liposomal paclitaxel, radiation or liposomal paclitaxel plus radiation treatment in terms of cell viability, apoptosis, colony formation and morphological changes. Future studies are required to evaluate these effects in vivo.