The synergistic effect of KU0060648 and TMZ was also observed in the treatment of glioma in vivo. High dose of KU0060648 addition to TMZ had a more remarkable anti-tumor effect than the low dose of KU0060648 in the presence of TMZ as well as KU0060648 at its high dose alone, further suggesting the feasibility of combination of DNA-PKcs inhibitor and TMZ on the glioma growth. 100 μM TMZ alone could not upregulate the survival time in animal harboring U87 tumors. However, with increased dose, KU0060648 sufficiently extended the survival time of TMZ treatment group, confirming that the combination of KU0060648 and TMZ had a synergistic effect for prolonging the survival period of nude mice harboring U87 xenograft tumors. Collectively, these data expanded the feasibility of the combination of the DNA-PKcs inhibitor with chemotherapeutic agents in solid tumor treatment.