gms | German Medical Science

58. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie e. V. (DGNC)

Deutsche Gesellschaft für Neurochirurgie (DGNC) e. V.

26. bis 29.04.2007, Leipzig

Inhibition of malignant glioma cell growth by retroviral shRNA-targeting of chromosomal passenger proteins Survivin and INCENP

Gezielte Inaktivierung der "Chromosomal Passenger"-Proteine INCENP und Survivin mittels retroviraler shRNA-Vektoren zur experimentellen Gentherapie des Glioblastomas multiforme

Meeting Abstract

  • corresponding author S. Hendruschk - Klinik für Neurochirugie, Medizinische Fakultät Carl Gustav Carus der Technischen Universität Dresden, Dresden, Deutschland
  • E. P. Rieber - Institut für Immunologie, Medizinische Fakultät Carl Gustav Carus der Technischen Universität Dresden, Dresden, Deutschland
  • G. Schackert - Klinik für Neurochirugie, Medizinische Fakultät Carl Gustav Carus der Technischen Universität Dresden, Dresden, Deutschland
  • A. Temme - Klinik für Neurochirugie, Medizinische Fakultät Carl Gustav Carus der Technischen Universität Dresden, Dresden, Deutschland

Deutsche Gesellschaft für Neurochirurgie. 58. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie e.V. (DGNC). Leipzig, 26.-29.04.2007. Düsseldorf: German Medical Science GMS Publishing House; 2007. DocP 072

Die elektronische Version dieses Artikels ist vollständig und ist verfügbar unter: http://www.egms.de/de/meetings/dgnc2007/07dgnc327.shtml

Veröffentlicht: 11. April 2007

© 2007 Hendruschk et al.
Dieser Artikel ist ein Open Access-Artikel und steht unter den Creative Commons Lizenzbedingungen (http://creativecommons.org/licenses/by-nc-nd/3.0/deed.de). Er darf vervielf&aauml;ltigt, verbreitet und &oauml;ffentlich zug&aauml;nglich gemacht werden, vorausgesetzt dass Autor und Quelle genannt werden.


Gliederung

Text

Objective: Glioblastoma multiforme, a malignant brain tumor, exhibits dysregulated apoptosis and enhanced cell growth due to genetic alterations and is resistant to conventional radiotherapy and chemotherapy. Thus, it is necessary to establish new approaches for a successful treatment. A new strategy for the destruction of glioma cells is the specific silencing of effector proteins of mitosis. By retroviral gene delivery we sought to introduce shRNAs for the inhibition of the “Chromosomal Passenger” proteins (CPPs) INCENP (Inner Centromeric Protein) and Survivin into glioma cells. Silencing of these proteins interferes with the cell cycle and leads to p53-independent cell death.

Methods: A retroviral vector system for delivery of shRNAs into glioma cells was established. RNAi-mediated down-regulation of protein expression was proven by Western Blot analyses and confocal laser scanning microscopy. The effects caused by the loss of Survivin and INCENP on the cell cycle were investigated by flow cytometry. Long-term survival of GBM cells with inactivated “Chromosomal Passenger” proteins was visualized by Giemsa staining.

Results: Transduction of cells with shRNA against either Survivin or shINCENP resulted in polyploidy and growth inhibition of glioblastoma cells. The high degree of polyploidy seen early after transduction was followed by apoptosis as detected by internucleosomal DNA fragmentation analysis. In addition, clonogenic assays verified the strong cytotoxic effects mediated by the transduced shRNA against Survivin and INCENP, respectively.

Conclusions: The inactivation of CPPs leads to mitotic defects and subsequently to apoptosis in glioma cells. This suggests a p53-independent inhibitory effect of shSurvivin and shINCENP on these cells, which might be beneficial for the treatment of malignant brain tumors.