Abstract
Background
Treatments which significantly improve progression-free and overall survival for patients with relapsed glioblastoma (GBM) after the standard therapy are lacking. The Topoisomerase II (TopoII) enzyme is a key target of anticancer agents because of the important role it plays in transcription regulation and chromatin remodeling. A drug with strong topoisomerase-mediated anticancer activity is etoposide that is used in combination with carboplatin in patients with relapsed GBM. We hypothesized that tumors harboring high expression of TopoII alpha (TopoIIa) would be more sensitive to etoposide treatment.
Methods
The relative expression levels of TopoIIa protein were measured in a panel of GBM cell lines using Western blot analysis and in a cohort of GBM using immunohistochemistry. Expression levels of TopoIIa in the cell lines were correlated with relative sensitivity to treatment with etoposide. To ascertain the role TopoIIa plays in mediating response to etoposide, expression was reduced with a siRNA targeted to TopoIIa.
Results
Protein expression of TopoIIa, although high in the cell lines, was very low in patient specimens. Correlations between TopoIIa protein expression and sensitivity to etoposide were evident. The IC50 for the low-TopoIIa-expressing cell line, T98G, was almost 50 times higher than M059K (high TopoIIa). Inhibition of TopoIIa in MO59K cells with siRNA significantly altered the IC50, increasing the resistance to etoposide. Interestingly, the expression of TopoIIa was not decreased after treatment with etoposide, indicating other mechanisms underplay treatment response.
Conclusions
In vitro, the levels of TopoIIa protein expression correlate with response to etoposide but also multiple molecular events namely DNA-PK and MDR also play a role in cell sensitivity to etoposide. That we did not find a high expression of TopoIIa in clinical specimens further suggests the mechanisms underlying treatment response are complex.
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Acknowledgments
The authors would like to acknowledge the assistance of Ms Sanaz Maleki for help with the immunohistochemistry and Dr Helen Wheeler for assisting the clinical data collection. HS was funded by the Postgraduate Research Scholarship from The Ministry of National Education, Republic of Turkey and consumables for this project funded by the Andrew Olle Memorial Trust and the Sydney Neuro-oncology Group.
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Sevim, H., Parkinson, J.F. & McDonald, K.L. Etoposide-mediated glioblastoma cell death: dependent or independent on the expression of its target, topoisomerase II alpha?. J Cancer Res Clin Oncol 137, 1705–1712 (2011). https://doi.org/10.1007/s00432-011-1046-5
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DOI: https://doi.org/10.1007/s00432-011-1046-5