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Ku70 and Rad51 vary in their importance for the repair of doxorubicin- versus etoposide-induced DNA damage

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Abstract

For DNA targeting anticancer drugs, cellular DNA repair mechanisms may cause resistance and hamper the therapeutic outcome. DNA damage induced by topoisomerase IIα inhibitors like etoposide and anthracyclines, which are a mainstay of cancer therapy, is also repaired in many cell types, but the impact and precise mechanisms of this repair are still obscure. To investigate the DNA damage response of human adenocarcinoma HT29-cells to doxorubicin and to compare the involvement of Ku70 and Rad51 in the repair of doxorubicin- versus etoposide-induced DNA damage, we assessed cell cycle distribution and cell death, DNA damage, proteins relevant for repair by homologous recombination and non-homologous end-joining, and clonogenicity following exposure to doxorubicin at clinically achievable concentrations. Also, we assessed changes in the repair kinetics after siRNA-mediated attenuation of Ku70 or Rad51 expression. We found that exposure to doxorubicin for 24 h induced a substantial amount of DNA damage that was largely repaired when doxorubicin was removed and the cells were maintained in drug-free medium. Nevertheless, a pronounced G2/M arrest occurred at times when repair was maximal. This was followed by a distinct increase in cell death and loss of clonogenicity. In this regard, responses to doxorubicin and etoposide were similar. However, distinct differences in the repair process following doxorubicin versus etoposide were seen in concentration dependency, time-course and requirement of Ku70 and Rad51 proteins. In spite of the shared molecular target of doxorubicin and etoposide, DNA lesions induced by these compounds are repaired differently.

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Acknowledgments

We would like to thank Prof. Dr. Frank Gieseler from the University Hospital Luebeck for many helpful discussions. Our work was supported by the Ernst und Elfriede Griebel’s Foerderungs- und Unterstuetzungsstiftung, Hamburg.

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The authors declare that they have no conflict of interest.

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  1. Clinical Pharmacy, Institute of Pharmacy, Hamburg University, Bundesstr. 45, 20146, Hamburg, Germany

    Ilona Schonn, Jana Hennesen & Dorothee C. Dartsch

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  1. Ilona Schonn
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  2. Jana Hennesen
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  3. Dorothee C. Dartsch
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Correspondence to Dorothee C. Dartsch.

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Schonn, I., Hennesen, J. & Dartsch, D.C. Ku70 and Rad51 vary in their importance for the repair of doxorubicin- versus etoposide-induced DNA damage. Apoptosis 16, 359–369 (2011). https://doi.org/10.1007/s10495-010-0564-y

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