Abstract
Purpose
Patients with oral squamous cell carcinomas (OSCC) often receive radiotherapy to preferentially induce apoptosis of cancer cells through generation of overwhelming DNA damage. This is amplified by generation of reactive oxygen species (ROS), thereby causing oxidative stress and cell death. However, tumors resist through different mechanisms, including upregulation of anti-apoptotic factors and enhanced ROS resistance. We recently reported that the antioxidative enzyme PON2 significantly enhances cellular stress resistance by attenuating mitochondrial ROS-mediated apoptosis. Further, PON2 is often upregulated in cancer. This prompted us to investigate its yet unknown role in the protection of OSCC against irradiation-induced cell death.
Methods
PON2 expression was determined after 7 Gy singular irradiation in four OSCC cell lines (PCI-13, PCI-52, SCC-4, SCC-68) accompanied by the detection of caspase 3/7 activity. A direct role of PON2 was tested by siRNA-mediated knockdown. In vivo PON2 expression was tested in five patients with oral carcinoma and compared with healthy mucosa for the evaluation of clinical significance.
Results
PON2 is variably expressed in OSCC in vitro and in vivo. Compared with the other cell lines, SCC-4 cells showed twofold more basal PON2 (p ≤ 0.05) and the lowest caspase 3/7 activity after singular irradiation (p ≤ 0.05). Contrarily, irradiation led to 1.2-fold induction of PON2 in PCI-13 with no effect on SCC-4 (≤0.05), suggesting that PON2 levels reflect the cells’ irradiation sensitivity. In agreement, PON2 knockdown resulted in significant higher apoptosis rates (p ≤ 0.05).
Conclusion
Our findings give first evidence that upregulation of PON2 may protect OSCC against irradiation-induced apoptosis.
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Acknowledgments
Thanks to Jutta Bühler, Jutta Goldschmitt, Lotte Groothusen, Ute Zerfaß and Petra Wilgenbus for their excellent technical assistance. Maximilian Krüger has been financially supported by internal funds of the University Medical Center Mainz (MAIFOR) and the German Research Foundation (DFG-Grant KR 4393/1-1). Sven Horke was funded by a U.S. Department of Defense award (W81XWH-12-2-0091), intramural funds from the University Medical Center Mainz, the German Research Foundation (DO1289/6-1), Gerhard and Martha Röttger foundation and the Center for Thrombosis and Hemostasis, Mainz (BMBF funding allocation ID 01E01003, project TRPA11i).
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The authors declare that they have no conflict of interest. All authors read and approved the final manuscript.
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All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. This article does not contain any studies with animals performed by any of the authors.
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The authors declare that parts of the results of the study were only presented in the form of preliminary result presentation as poster presentation on the 30th German Cancer Congress 2012 (DKK) and as poster supplement of the 64th congress of the DGMKG (Deutsche Gesellschaft für Mund-, Kiefer-, und Gesichtschirurgie) in the International Poster Journal of Dentistry and Oral medicine, but not in the same form as presented here.
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Krüger, M., Pabst, A.M., Al-Nawas, B. et al. Paraoxonase-2 (PON2) protects oral squamous cell cancer cells against irradiation-induced apoptosis. J Cancer Res Clin Oncol 141, 1757–1766 (2015). https://doi.org/10.1007/s00432-015-1941-2
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DOI: https://doi.org/10.1007/s00432-015-1941-2