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A novel Chk inhibitor, XL-844, increases human cancer cell radiosensitivity through promotion of mitotic catastrophe

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Summary

Check point kinases (Chk) play a major role in facilitating DNA repair upon radiation exposure. We tested the potency of a novel inhibitor of Chk1 and Chk2, XL-844 (provided by Exelixis Inc., CA, USA), to radiosensitize human cancer cells grown in culture and investigated the underlying mechanisms. HT-29 cells (a human colon cancer line) were exposed to XL-844, radiation, or both, and assessed for clonogenic cell survival. Treatment-dependent effects on phosphorylated forms of Chk proteins were assessed by Western blots. Further mechanistic investigations in HT-29 cells included cell cycle analysis by flowcytometry and assessment of DNA repair kinetics by immuno-cytochemistry (ICC) for nuclear appearance of the phosphorylated form of histone 2AX protein (γ-H2AX) staining. Cells undergoing mitotic catastrophe were identified by irregular pattern of mitotic spindle markers α and γ-tubulin staining by ICC. XL-844 enhanced radiosensitivity in a dose and schedule-dependent manner and the enhancement factor was 1.42 at 0.5 survival fraction. Mechanistically XL-844 abrogated radiation-induced Chk2 phosphorylation, induced pan-nuclear γ-H2AX, and prolonged the presence of radiation-induced γ-H2AX foci, and promoted mitotic catastrophe. In conclusion, our data showed that inhibition of Chk2 activity by XL-844 enhanced cancer cell radiosensitivity that was associated with inhibition of DNA repair and induction of mitotic catastrophe.

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Acknowledgements

This work was supported partly by Exelixis Inc., CA, USA (PI: U. Raju) and partly by a UICC ACSBI fellowship supplemented by the University of Zurich (O. Riesterer). We thank Peter Lamb Ph.D. (Exelixis Incorporation, San Francisco, CA) for his involvement in designing the experiments.

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  1. Oliver Riesterer

    Present address: Department of Radiation Oncology, University Hospital Zurich, Ramistrasse 100, 8091, Zurich, Switzerland

Authors and Affiliations

  1. Department of Experimental Radiation Oncology, The University of Texas M. D. Anderson Cancer Center, Unit 66, 1515 Holcombe Blvd., Houston, TX, 77030, USA

    Oliver Riesterer, Fumihiko Matsumoto, Li Wang, Jessica Pickett, David Molkentine, Uma Giri, Luka Milas & Uma Raju

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  1. Oliver Riesterer
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Correspondence to Uma Raju.

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Riesterer, O., Matsumoto, F., Wang, L. et al. A novel Chk inhibitor, XL-844, increases human cancer cell radiosensitivity through promotion of mitotic catastrophe. Invest New Drugs 29, 514–522 (2011). https://doi.org/10.1007/s10637-009-9361-2

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