DNA-PKCs deficiency leads to persistence of oxidatively induced DNA lesions in human tumor cells

Prakash Peddi, Charles W Loftin, Jennifer S Dickey, Jessica M Hair, Kara J Burns, Khaled Aziz, Dave C Francisco, Mihalis I Panayiotidis, Olga A Sedelnikova, William M Bonner, Thomas A Winters, Alexandros G Georgakilas

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    Abstract

    DNA-dependent protein kinase (DNA-PK) is a key non-homologous-end-joining (NHEJ) nuclear serine/threonine protein kinase involved in various DNA metabolic and damage signaling pathways contributing to the maintenance of genomic stability and prevention of cancer. To examine the role of DNA-PK in processing of non-DSB clustered DNA damage, we have used three models of DNA-PK deficiency, i.e., chemical inactivation of its kinase activity by the novel inhibitors IC86621 and NU7026, knockdown and complete absence of the protein in human breast cancer (MCF-7) and glioblastoma cell lines (MO59-J/K). A compromised DNA-PK repair pathway led to the accumulation of clustered DNA lesions induced by gamma-rays. Tumor cells lacking protein expression or with inhibited kinase activity showed a marked decrease in their ability to process oxidatively induced non-DSB clustered DNA lesions measured using a modified version of pulsed-field gel electrophoresis or single-cell gel electrophoresis (comet assay). In all cases, DNA-PK inactivation led to a higher level of lesion persistence even after 24-72h of repair. We suggest a model in which DNA-PK deficiency affects the processing of these clusters first by compromising base excision repair and second by the presence of catalytically inactive DNA-PK inhibiting the efficient processing of these lesions owing to the failure of DNA-PK to disassociate from the DNA ends. The information rendered will be important for understanding not only cancer etiology in the presence of an NHEJ deficiency but also cancer treatments based on the induction of oxidative stress and inhibition of cluster repair.
    Original languageEnglish
    Pages (from-to)1435-1443
    Number of pages8
    JournalFree Radical Biology and Medicine
    Volume48
    Issue number10
    DOIs
    Publication statusPublished - 15 May 2010

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  • Cite this

    Peddi, P., Loftin, C. W., Dickey, J. S., Hair, J. M., Burns, K. J., Aziz, K., Francisco, D. C., Panayiotidis, M. I., Sedelnikova, O. A., Bonner, W. M., Winters, T. A., & Georgakilas, A. G. (2010). DNA-PKCs deficiency leads to persistence of oxidatively induced DNA lesions in human tumor cells. Free Radical Biology and Medicine, 48(10), 1435-1443. https://doi.org/10.1016/j.freeradbiomed.2010.02.033