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  • Original Article
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Knockdown of survivin expression by small interfering RNA reduces the clonogenic survival of human sarcoma cell lines independently of p53

Cancer Gene Therapy volume 11, pages 186–193 (2004)Cite this article

Abstract

Survivin, a member of the inhibitors-of-apoptosis gene family, is overexpressed in many tumor types. Survivin is a prognostic marker of soft-tissue sarcomas, but the downregulation of survivin expression and the possible dependency of survivin downregulation on p53 in these tumors have not been investigated. Therefore, we applied small interfering RNA (siRNA) to knock down the expression of survivin in five human sarcoma cell lines with wild-type or mutant p53 alleles. Compared with survivin mRNA expression in the nonsense siRNA-treated sarcoma cell lines, expression after treatment with survivin-specific siRNA was reduced by 73–88%; survivin protein expression was reduced by 52–81%. This finding was coupled with a reduction in clonogenic survival ranging from 65–86%. However, less than 10% of cells treated with survivin-specific siRNA underwent apoptosis. Cell-cycle and morphologic analyses showed that after a dramatic increase in the number of treated cells in the G2/M phase, some of the cells became polyploid; this result indicates that mitosis of a substantial number of treated cells was incomplete. Our findings suggest that survivin-specific siRNA could be a selective treatment to kill sarcoma cells regardless of the presence or absence of wild-type p53 alleles.

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Acknowledgements

We thank our colleagues from the Institute of Pathology for contributing to this study and for their continuous support. We also thank Ute Rolle, Kathrin Spröte, and Sandy Kaufhold (Institute of Pathology, Martin-Luther-University Halle-Wittenberg), and Antje Zobjack (Institute of Pathology, Technical University Dresden) for their excellent technical assistance. We are very grateful to Uta Schramm (Department of Dermatology, Martin-Luther-University Halle-Wittenberg) for helping with the flow cytometric analyses and to Clare Burns-Klein for revising the manuscript. We are very thankful for the editing of the manuscript by Dr Julia Cay Jones (Department of Scientific Editing, St Jude Children's Research Hospital, Memphis, TN). This work was supported by the State Saxony-Anhalt (Grant Number: 3347A/0021B), the Wilhelm Roux-program of BMBF/NBL3 (FKZ: 4/18), by the Wilhelm Sander-Stiftung (Grant Number 99.009.1), and by ‘Förderverein GSGT e.V.’ Frank Bartel and Karen Blümke were supported by a grant from Chiron Behring GmbH (Marburg, Germany) and Frank Bartel by the Deutsche Krebshilfe, Mildred Scheel Shiftung Fuz no-2130-Ta2.

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Author notes

  1. Helge Taubert

    Present address: Hartwell Center for Bioinformatics and Biotechnology, St Jude Children's Research Hospital, Memphis, TN, 38105-2794, USA

  2. Matthias Kappler and Matthias Bache: Both authors contributed equally to the study.

Authors and Affiliations

  1. Institute of Pathology, Martin-Luther-University Halle-Wittenberg, Halle(S), Germany

    Matthias Kappler, Frank Bartel, Matti Panian, Hannelore Schmidt & Helge Taubert

  2. Department of Radiotherapy, Martin-Luther-University Halle-Wittenberg, Halle(S), Germany

    Matthias Bache

  3. Department of Urology, Martin-Luther-University Halle-Wittenberg, Halle(S), Germany

    Frank Bartel & Karen Blümke

  4. Institute of Pathology, Technical University Dresden, Dresden, Germany

    Matthias Kotzsch

  5. Clinic of General and Transplantation Surgery, University of Ulm, Ulm, Germany

    Peter Würl

  6. Department of Urology, Technical University Dresden, Dresden, Germany

    Axel Meye

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Kappler, M., Bache, M., Bartel, F. et al. Knockdown of survivin expression by small interfering RNA reduces the clonogenic survival of human sarcoma cell lines independently of p53. Cancer Gene Ther 11, 186–193 (2004). https://doi.org/10.1038/sj.cgt.7700677

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