We next determined the effect of the combination of KU0060648 and TMZ on the regulation of downstream effectors of AKT signaling and malignant markers in tumorous tissues. IHC analysis revealed that KU0060648 or KU0060648/TMZ treatment effectively inhibited the expression of p-DNA-PKcs and p-AKT (Figure 7C). In addition, staining of γH2AX was surveyed. Consistent with our in vitro study, significant increases of γH2AX levels only emerged in comparison between KU0060648/TMZ and TMZ groups, while not in KU0060648 treatment alone. KU0060648 or KU0060648/TMZ treatment also significantly suppressed the expression of Ki-67 (the biomarker of proliferation), c-Myc, survivin, MMP-9 and VEGF and enhanced the expression of apoptotic promoting biomarker caspase-3 in U87 tumors (Figure 7C). Collectively, these results suggested that combination of KU0060648 and TMZ was more efficient in inhibition of the U87 tumors than either of the single agents.