Investigational New Drugs

, Volume 29, Issue 3, pp 514–522 | Cite as

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

  • Oliver Riesterer
  • Fumihiko Matsumoto
  • Li Wang
  • Jessica Pickett
  • David Molkentine
  • Uma Giri
  • Luka Milas
  • Uma Raju
SHORT REPORT

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.

Keywords

Radiosensitivity Inhibitor of Check point kinases XL-844 Mitotic catastrophe Pan-nuclear γ-H2AX 

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Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Oliver Riesterer
    • 1
    • 2
  • Fumihiko Matsumoto
    • 1
  • Li Wang
    • 1
  • Jessica Pickett
    • 1
  • David Molkentine
    • 1
  • Uma Giri
    • 1
  • Luka Milas
    • 1
  • Uma Raju
    • 1
  1. 1.Department of Experimental Radiation OncologyThe University of Texas M. D. Anderson Cancer CenterHoustonUSA
  2. 2.Department of Radiation OncologyUniversity Hospital ZurichZurichSwitzerland

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