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Astrocytoma is the most common primary tumor of the brain. High-grade astrocytomas are aggressive tumors with poor prognosis. Biomarkers that predict patient outcome and response to a specific treatment will provide patients with personalized estimation of prognosis and customized therapeutic regimens with maximal effectiveness and minimal toxicity.The aim of this work was to explore the ability of the expression of IDH1 R132H, pSTAT3, and Tbx2 proteins to predict outcome of patients with high-grade astrocytoma.The expression of IDH1 R132H, pSTAT3, and Tbx2 proteins was assessed by immunohistochemistry in 41 high-grade astrocytoma cases. They were correlated with patients’ clinicopathological features, recurrence-free survival, overall survival (OS), and response to therapy.A significant direct association was observed between IDH1 R132H immunoreactivity and response to TMZ therapy (P=0.01), better progression-free survival (PFS) as well as OS (P<0.001). The expression of both Tbx2 and pSTAT3 proteins was inversely correlated with response to TMZ therapy (P=0.019 and 0.014, respectively). Survival analysis revealed that Tbx2 and pSTAT3 immunoreactivity was significantly associated with shorter PFS (P=0.021 and 0.047, respectively), as well as poor OS (P=0.005 and 0.007, respectively).IDH1 R132H protein expression and loss of Tbx2 and pSTAT3 proteins are associated with prolonged PFS and OS and better response to TMZ chemotherapy.