Hausmann Christina, Temme Achim, Cordes Nils, Eke Iris
OncoRay - National Center for Radiation Research in Oncology, Medical Faculty Carl Gustav Carus, Technische Universität Dresden, 01307 Dresden, Germany.
Section of Experimental Neurosurgery/Tumor Immunology, Department of Neurosurgery University Hospital Carl Gustav Carus, Technische Universität Dresden, 01307 Dresden, Germany.
Oncotarget. 2015 Oct 27;6(33):34592-605. doi: 10.18632/oncotarget.5423.
The prognosis is generally poor for patients suffering from glioblastoma multiforme (GBM) due to radiation and drug resistance. Prosurvival signaling originating from focal adhesion hubs essentially contributes to therapy resistance and tumor aggressiveness. As the underlying molecular mechanisms remain largely elusive, we addressed whether targeting of the focal adhesion proteins particularly interesting new cysteine-histidine-rich 1 (PINCH1), integrin-linked kinase (ILK) and ILK associated phosphatase (ILKAP) modulates GBM cell radioresistance. Intriguingly, PINCH1, ILK and ILKAP depletion sensitized p53-wildtype, but not p53-mutant, GBM cells to radiotherapy. Concomitantly, these cells showed inactivated Glycogen synthase kinase-3β (GSK3β) and reduced proliferation. For PINCH1 and ILKAP knockdown, elevated levels of radiation-induced γH2AX/53BP1-positive foci, as a marker for DNA double strand breaks, were observed. Mechanistically, we identified radiation-induced phosphorylation of DNA protein kinase (DNAPK), an important DNA repair protein, to be dependent on ILKAP. This interaction was fundamental to radiation survival of p53-wildtype GBM cells. Conclusively, our data suggest an essential role of PINCH1, ILK and ILKAP for the radioresistance of p53-wildtype GBM cells and provide evidence for DNAPK functioning as a central mediator of ILKAP signaling. Strategies for targeting focal adhesion proteins in combination with radiotherapy might be a promising approach for patients with GBM.
由于辐射和耐药性,多形性胶质母细胞瘤(GBM)患者的预后通常较差。源自粘着斑枢纽的促生存信号在很大程度上导致了治疗耐药性和肿瘤侵袭性。由于潜在的分子机制仍不清楚,我们研究了靶向粘着斑蛋白,特别是富含半胱氨酸和组氨酸的新蛋白1(PINCH1)、整合素连接激酶(ILK)和ILK相关磷酸酶(ILKAP)是否能调节GBM细胞的放射抗性。有趣的是,PINCH1、ILK和ILKAP的缺失使p53野生型而非p53突变型GBM细胞对放疗敏感。同时,这些细胞显示糖原合酶激酶-3β(GSK3β)失活且增殖减少。对于PINCH1和ILKAP基因敲低,观察到辐射诱导的γH2AX/53BP1阳性灶水平升高,γH2AX/53BP1是DNA双链断裂的标志物。从机制上讲,我们发现重要的DNA修复蛋白DNA蛋白激酶(DNAPK)的辐射诱导磷酸化依赖于ILKAP。这种相互作用对于p53野生型GBM细胞的辐射存活至关重要。总之,我们的数据表明PINCH1、ILK和ILKAP在p53野生型GBM细胞的放射抗性中起重要作用,并为DNAPK作为ILKAP信号的中心介质发挥作用提供了证据。针对粘着斑蛋白与放疗联合的策略可能是GBM患者的一种有前景的方法。