Glioblastoma multiforme (GBM) is a highly malignant and notably aggressive primary tumour. Variant III of the epidermal growth factor receptor (EGFRvIII) is one of the most common types of variants in GBM, and serves an important role in tumour invasion, proliferation and treatment resistance. In the present study, statistical analyses were performed on data from 57 patients with GBM, and polymerase chain reaction detection was conducted on the tumour tissues from 32 of these patients. The results indicated that the EGFRvIII mutation was significantly associated with tumour malignancy. Human GBMU87-EGFRvIII cell lines were cultured and treated with radiosurgery and temozolomide individually, or with combined radiosurgery and temozolomide treatment. In vitro and in vivo experimental methods were used to detect the expression levels of Ki-67 and EGFRvIII. As verified in the present study, the EGFRvIII mutation is positively correlated with the malignancy of tumours, and combined radiosurgery and temozolomide therapy may inhibit the invasion and proliferation abilities of U87-EGFRvIII more effectively than treatment alone.