Preclinical Development NF-kB Is Required for SmacMimetic-Mediated Sensitization of Glioblastoma Cells for g-Irradiation–Induced Apoptosis

Evasion of apoptosis contributes to radioresistance of glioblastoma, calling for novel strategies to overcome apoptosis resistance. In this study, we investigated the potential of the small molecule Smac mimetic BV6 to modulate radiosensitivity of glioblastoma cells. Here, we identify a novel proapoptotic function of NF-kB in g-irradiation–induced apoptosis of glioblastoma cells by showing, for the first time, that NF-kB is critically required for Smac mimetic–mediated radiosensitization. BV6 significantly increases g-irradiation–triggered apoptosis in several glioblastoma cell lines in a doseand time-dependentmanner. Calculation of combination index (CI) reveals that the interaction of BV6 and g-irradiation is highly synergistic (CI < 0.3).Molecular studies show that BV6 stimulates NF-kB activation, which is critical for radiosensitization, because genetic inhibition of NF-kB by overexpression of the dominant-negative superrepressor IkBa-SR significantly decreases BV6and g-irradiation–induced apoptosis. Also, the BV6-mediated enhancement of g-irradiation–triggered caspase activation, drop of mitochondrial membrane potential, and cytochrome c release is abolished in cells overexpressing IkBa-SR. Similarly, NF-kB inhibition by ectopic expression of a kinase dead mutant of IKKb prevents the BV6-mediated sensitization for g-irradiation. The clinical relevance is underscored by experiments with primary tumor samples showing that BV6 sensitizes primary cultured glioma cells as well as glioblastoma-initiating cancer stem cells derived from surgical specimens for g-irradiation. In conclusion, we identify NF-kB as a critical mediator of Smac mimetic-conferred radiosensitization of glioblastoma cells. These results have important implications for the development of Smacmimetic–based combination protocols for radiosensitization of glioblastoma. Mol Cancer Ther; 10(10); 1867–75. 2011 AACR.