Analyzing the influence of kinase inhibitors on DNA repair by differential proteomics of chromatin-interacting proteins and nuclear phospho-proteins
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Lisa Gleißner1,*, Marcel Kwiatkowski2,3,*, Laura Myllynen1, Pascal Steffen2, Cordula Petersen1, Kai Rothkamm1, Hartmut Schlüter2,* and Malte Kriegs1,*
1Laboratory of Radiobiology and Experimental Radiation Oncology, Hubertus Wald Tumorzentrum, University Cancer Center Hamburg, University Medical Center Hamburg, Eppendorf, Hamburg, Germany
2Institute of Clinical Chemistry, Hubertus Wald Tumorzentrum, University Cancer Center Hamburg, University Medical Center Hamburg, Eppendorf, Hamburg, Germany
3Department of Pharmacokinetics, Toxicology and Targeting, Groningen Research Institute for Pharmacy, University of Groningen, Groningen, The Netherlands
*These authors contributed equally to this work
Malte Kriegs, email: firstname.lastname@example.org
Keywords: differential proteomics; phospho-proteins; chromatin bound proteins; sorafenib; head and neck cancer
Received: August 18, 2017 Accepted: October 25, 2017 Published: November 10, 2017
The combination of radiotherapy and pharmacological inhibition of cellular signal transduction pathways offers promising strategies for enhanced cancer cell inactivation. However, the molecular effects of kinase inhibitors especially on DNA damage detection and repair after X-irradiation have to be understood to facilitate the development of efficient and personalized treatment regimens. Therefore, we applied differential proteomics for analyzing inhibitor-induced changes in either chromatin-bound or phosphorylated nuclear proteins. The effect of the multi kinase inhibitor sorafenib on DNA repair, chromatin binding and phosphorylation of nuclear proteins was analyzed in UT-SCC 42B head and neck cancer cells using metabolic labeling based differential proteomics (SILAC). Sorafenib significantly inhibited DNA repair but failed to significantly affect chromatin interactions of 90 quantified proteins. In contrast, analyzing nuclear phospho-proteins following sorafenib treatment, we detected quantitative changes in 9 out of 59 proteins, including DNA-repair proteins. In conclusion, the analysis of nuclear phospho-proteins by differential proteomics is an effective tool for determining the molecular effects of kinase inhibitors on X-irradiated cells. Analyzing chromatin binding might be less promising.
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