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The effects of knockdown of wild-type survivin, survivin-2B or survivin-Δ3 on the radiosensitization in a soft tissue sarcoma cells in vitro under different oxygen conditions

Cancer Gene Therapy volume 14, pages 994–1001 (2007)Cite this article

Abstract

The inhibitor of apoptosis wild-type survivin is a multifunctional protein that suppresses apoptosis and regulates cell cycle progression. An association between wild-type survivin expression and radiosensitivity has been described in different tumor cells. The effects of siRNA-induced knockdown of wild-type survivin and survivin-splice variants survivin-2B and survivin-Δ3 were investigated under normoxic and hypoxic conditions in the human sarcoma cell line US 8–93 (mutant p53). Inhibition of the survivin isoforms by siRNA resulted in a decrease of target mRNA down to 14–70% compared to cells treated with control siRNA independent of the oxygen level. The mRNA expression of survivin isoforms was decreased by the factor of 1–12 when the cells were cultivated under hypoxic conditions. Moreover, the knockdown of wild-type survivin reduced colony formation independent of oxygen concentration down to 70% and induced formation of polyploid cells. Less reduction of plating efficiency was observed after specific knockdown of survivin-2B and survivin-Δ3 under hypoxic or normoxic conditions. A knockdown of wild-type survivin, survivin-Δ3 and survivin-2B isoforms in combination with irradiation caused no radiosensitization in cell line US 8–93, neither under hypoxic nor under normoxic conditions tested in the colony-forming assay. However, knockdown of wild-type survivin caused radiosensitization in the megacolony assay.

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Abbreviations

siRNA:

small interfering RNA

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Acknowledgements

We thank our colleagues from the Department of Radiotherapy and the Institute of Pathology for contributing to this study and for their continuous support. We also thank Kathrin Spröte and Ute Rolle (Department of Radiotherapy, Martin-Luther-University Halle-Wittenberg) for their excellent technical assistance. We are very grateful to Uta Schramm und Dr Thomas Klapperstück (Department of Dermatology, Martin-Luther-University Halle-Wittenberg) for helping at the flow cytometric analyses. Furthermore, we thank Dr Axel Meye for helpful discussion. Major parts of the results were achieved during participation in the MSc course in Radiation Biology of University of College London (2005–2006) organized by Professor Klaus-R Trott and Dr Kevin M Prise. This work was supported by the State of Saxony-Anhalt (Grant number: 3584AB/1104 M) and by the Wilhelm Roux-Program of BMBF/NBL3 (FKZ: 16/18 and FKZ: 15/18).

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Author notes

  1. M Kappler and S Rot: These authors have contributed equally to this work.

Authors and Affiliations

  1. Department of Radiotherapy, Faculty of Medicine, University of Halle-Wittenberg, Halle/Saale, Germany

    M Kappler, K Dellas, G Hänsgen & M Bache

  2. Institute of Pathology, University of Halle-Wittenberg, Halle/Saale, Germany

    S Rot, H Taubert, T Greither & F Bartel

  3. Gray Cancer Institute, Mount Vernon Hospital, Northwood, UK

    K-R Trott

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  1. M Kappler
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Correspondence to M Kappler.

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Kappler, M., Rot, S., Taubert, H. et al. The effects of knockdown of wild-type survivin, survivin-2B or survivin-Δ3 on the radiosensitization in a soft tissue sarcoma cells in vitro under different oxygen conditions. Cancer Gene Ther 14, 994–1001 (2007). https://doi.org/10.1038/sj.cgt.7701090

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