Skip to main content
SpringerLink
Log in

Higher topoisomerase 2 alpha gene transcript levels predict better prognosis in GBM patients receiving temozolomide chemotherapy: identification of temozolomide as a TOP2A inhibitor

  • Laboratory Investigation - Human/Animal Tissue
  • Published:
Journal of Neuro-Oncology Aims and scope Submit manuscript

Abstract

The search for molecular markers which predict response to chemotherapy is an important aspect of current neuro-oncology research. MGMT promoter methylation is the only proved marker of glioblastoma. The purpose of this study was to assess the effect of topoisomerase expression on glioblastoma survival and study the mechanisms involved. The transcript levels of all isoforms of the topoisomerase family in all grades of diffuse astrocytoma were assessed. A prospective study of patients with glioblastoma treated by a uniform treatment procedure was performed with the objective of correlating outcome with gene expression. The ability of TOP2A enzyme to relax the super coiled plasmid DNA in the presence of temozolomide was evaluated to assess its effect on TOP2A. The temozolomide cyctotoxicity of TOP2A-silenced U251 cells was assessed. The transcript levels of TOP2A, TOP2B, and TOP3A are upregulated significantly in GBM in comparison with lower grades of astrocytoma and normal brain samples. mRNA levels of TOP2A correlated significantly with survival of the patients. Higher TOP2A transcript levels in GBM patients predicted better prognosis (P = 0.043; HR = 0.889). Interestingly, we noted that temozolomide inhibited TOP2A activity in in-vitro enzyme assays. We also noted that siRNA knock down of TOP2A rendered a glioma cell line resistant to temozolomide chemotherapy. We demonstrated for the first time that temozolomide is also a TOP2A inhibitor and established that TOP2A transcript levels determine the chemosensitivity of glioblastoma to temozolomide therapy. Very high levels of TOP2A are a good prognostic indicator in GBM patients receiving temozolomide chemotherapy.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

GBM:

Glioblastoma

DA:

Diffuse astrocytoma

AA:

Anaplastic astrocytoma

TOP2A:

Topoisomerase 2 alpha

References

  1. Furnari FB, Fenton T, Bachoo RM et al (2007) Malignant astrocytic glioma: genetics, biology, and paths to treatment. Genes Dev 21:2683–2710

    Article  PubMed  CAS  Google Scholar 

  2. Stewart LA (2002) Chemotherapy in adult high-grade glioma: a systematic review and meta-analysis of individual patient data from 12 randomised trials. Lancet 359:1011–1018

    Article  PubMed  CAS  Google Scholar 

  3. Stupp R, Hegi ME, Mason WP et al (2009) Effects of radiotherapy with concomitant and adjuvant temozolomide versus radiotherapy alone on survival in glioblastoma in a randomised phase III study: 5-year analysis of the EORTC-NCIC trial. Lancet Oncol 10:459–466

    Article  PubMed  CAS  Google Scholar 

  4. Stevnsner T, Bohr VA (1993) Studies on the role of topoisomerases in general, gene- and strand-specific DNA repair. Carcinogenesis 14:1841–1850

    Article  PubMed  CAS  Google Scholar 

  5. Watt PM, Hickson ID (1994) Structure and function of type II DNA topoisomerases. Biochem J 303(Pt 3):681–695

    PubMed  CAS  Google Scholar 

  6. Kasahara K, Fujiwara Y, Sugimoto Y et al (1992) Determinants of response to the DNA topoisomerase II inhibitors doxorubicin and etoposide in human lung cancer cell lines. J Natl Cancer Inst 84:113–118

    Article  PubMed  CAS  Google Scholar 

  7. Shpitz B, Bomstein Y, Zehavi T et al (2000) Topoisomerase II alpha expression in ductal carcinoma in situ of the breast: a preliminary study. Hum Pathol 31:1249–1254

    Article  PubMed  CAS  Google Scholar 

  8. Taniguchi K, Wakabayashi T, Yoshida T et al (1999) Immunohistochemical staining of DNA topoisomerase II alpha in human gliomas. J Neurosurg 91:477–482

    Article  PubMed  CAS  Google Scholar 

  9. Hao YJ, Natsume A, Mizuno M et al (2000) Correlation between DNA topoisomerase II alpha expression and sensitivity to etoposide in human glioma cell lines. Gan To Kagaku Ryoho 27:1403–1409

    PubMed  CAS  Google Scholar 

  10. Hashimoto S, Chatterjee S, Ranjit GB et al (1995) Drastic reduction of topoisomerase II alpha associated with major acquired resistance to topoisomerase II active agents but minor perturbations of cell growth. Oncol Res 7:407–416

    PubMed  CAS  Google Scholar 

  11. Asano T, An T, Mayes J et al (1996) Transfection of human topoisomerase II alpha into etoposide-resistant cells: transient increase in sensitivity followed by down-regulation of the endogenous gene. Biochem J 319(Pt 1):307–313

    PubMed  CAS  Google Scholar 

  12. Zhou Z, Zwelling LA, Ganapathi R, Kleinerman ES (2001) Enhanced etoposide sensitivity following adenovirus-mediated human topoisomerase II alpha gene transfer is independent of topoisomerase II beta. Br J Cancer 85:747–751

    Article  PubMed  CAS  Google Scholar 

  13. Zhou Z, Zwelling LA, Kawakami Y et al (1999) Adenovirus-mediated human topoisomerase II alpha gene transfer increases the sensitivity of etoposide-resistant human breast cancer cells. Cancer Res 59:4618–4624

    PubMed  CAS  Google Scholar 

  14. Gobble RM, Qin LX, Brill ER et al (2011) Expression profiling of liposarcoma yields a multigene predictor of patient outcome and identifies genes that contribute to liposarcomagenesis. Cancer Res 71:2697–2705

    Article  PubMed  CAS  Google Scholar 

  15. Hart MG, Grant R, Garside R et al. (2008) Temozolomide for high grade glioma. CocHRane Database Syst Rev; CD007415

  16. Hegi ME, Liu L, Herman JG et al (2008) Correlation of O6-methylguanine methyltransferase (MGMT) promoter methylation with clinical outcomes in glioblastoma and clinical strategies to modulate MGMT activity. J Clin Oncol 26:4189–4199

    Article  PubMed  CAS  Google Scholar 

  17. Mandraju RK, Kannapiran P, Kondapi AK (2008) Distinct roles of Topoisomerase II isoforms: DNA damage accelerating alpha, double strand break repair promoting beta. Arch Biochem Biophys 470:27–34

    Article  PubMed  CAS  Google Scholar 

  18. Jarvinen TA, Liu ET (2003) HER-2/neu and topoisomerase II alpha in breast cancer. Breast Cancer Res Treat 78:299–311

    Article  PubMed  Google Scholar 

  19. Wang JC (1996) DNA topoisomerases. Annu Rev Biochem 65:635–692

    Article  PubMed  CAS  Google Scholar 

  20. Isaacs RJ, Davies SL, Sandri MI et al (1998) Physiological regulation of eukaryotic topoisomerase II. Biochim Biophys Acta 1400:121–137

    PubMed  CAS  Google Scholar 

  21. Yang X, Li W, Prescott ED et al (2000) DNA topoisomerase II beta and neural development. Science 287:131–134

    Article  PubMed  CAS  Google Scholar 

  22. Oda M, Arakawa Y, Kano H et al (2005) Quantitative analysis of topoisomerase II alpha to rapidly evaluate cell proliferation in brain tumors. Biochem Biophys Res Commun 331:971–976

    Article  PubMed  CAS  Google Scholar 

  23. Zhao H, Yu H, Liu Y et al (2008) DNA topoisomerase II-alpha as a proliferation marker in human gliomas: correlation with PCNA expression and patient survival. Clin Neuropathol 27:83–90

    PubMed  CAS  Google Scholar 

  24. Ho DM, Hsu CY, Ting LT, Chiang H (2003) MIB-1 and DNA topoisomerase II alpha could be helpful for predicting long-term survival of patients with glioblastoma. Am J Clin Pathol 119:715–722

    Article  PubMed  CAS  Google Scholar 

  25. Faria MH, Goncalves BP, do Patrocinio RM et al (2006) Expression of Ki-67, topoisomerase IIalpha and c-MYC in astrocytic tumors: correlation with the histopathological grade and proliferative status. Neuropathology 26:519–527

    Article  PubMed  Google Scholar 

  26. Holden JA, Townsend JJ (1999) DNA topoisomerase II-alpha as a proliferation marker in astrocytic neoplasms of the central nervous system: correlation with MIB1 expression and patient survival. Mod Pathol 12:1094–1100

    PubMed  CAS  Google Scholar 

  27. Jarvinen TA, Tanner M, Rantanen V et al (2000) Amplification and deletion of topoisomerase II alpha associate with ErbB-2 amplification and affect sensitivity to topoisomerase II inhibitor doxorubicin in breast cancer. Am J Pathol 156:839–847

    Article  PubMed  CAS  Google Scholar 

  28. Gudkov AV, Zelnick CR, Kazarov AR et al (1993) Isolation of genetic suppressor elements, inducing resistance to topoisomerase II-interactive cytotoxic drugs, from human topoisomerase II cDNA. Proc Natl Acad Sci USA 90:3231–3235

    Article  PubMed  CAS  Google Scholar 

  29. Asano T, Zwelling LA, An T et al (1996) Effect of transfection of a Drosophila topoisomerase II gene into a human brain tumour cell line intrinsically resistant to etoposide. Br J Cancer 73:1373–1380

    Article  PubMed  CAS  Google Scholar 

  30. Vassetzky YS, Alghisi GC, Roberts E, Gasser SM (1996) Ectopic expression of inactive forms of yeast DNA topoisomerase II confers resistance to the anti-tumour drug, etoposide. Br J Cancer 73:1201–1209

    Article  PubMed  CAS  Google Scholar 

  31. Withoff S, Keith WN, Knol AJ et al (1996) Selection of a subpopulation with fewer DNA topoisomerase II alpha gene copies in a doxorubicin-resistant cell line panel. Br J Cancer 74:502–507

    Article  PubMed  CAS  Google Scholar 

  32. Yan L, Bulgar A, Miao Y, Mahajan V, Donze JR, Gerson SL, Liu L (2007) Combined treatment with temozolomide and methoxyamine: blocking apurininc/pyrimidinic site repair coupled with targeting topoisomerase IIα. Clin Cancer Res 13:1532–1539

    Article  PubMed  CAS  Google Scholar 

  33. Reardon DA, Quinn JA, Rich JN et al (2005) Phase I trial of irinotecan plus temozolomide in adults with recurrent malignant glioma. Cancer 104:1478–1486

    Article  PubMed  CAS  Google Scholar 

  34. Desjardins A, Reardon DA, Herndon JE et al (2008) Bevacizumab plus irinotecan in recurrent WHO grade 3 malignant gliomas. Clin Cancer Res 14:7068–7073

    Article  PubMed  CAS  Google Scholar 

  35. Leone S, Cornetta T, Basso E, Cozzi R (2010) Resveratrol induces DNA double-strand breaks tHRough human topoisomerase II interaction. Cancer Lett 295:167–172

    Article  PubMed  CAS  Google Scholar 

  36. Schmidt F, Knobbe CB, Frank B et al (2008) The topoisomerase II inhibitor, genistein, induces G2/M arrest and apoptosis in human malignant glioma cell lines. Oncol Rep 19:1061–1066

    PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This study was supported by a grant from the Council of Scientific and Industrial Research, Government of India, under the NMITLI program and Multi-institutional Network Programme in Molecular Neuro-oncology from DBT. We thank Shailaja Chandrashekararao, Cini Samuel, K Premkumar, and K Chandrasekar. The authors thank all the faculty from the Departments of Neurosurgery, NIMHANS and SSSIHMS who helped in conducting the study. Infrastructural support by funding from ICMR (Center for Advanced studies in Molecular Medicine), DBT, DST (FIST) and UGC to department of MCB, Indian Institute of Science is acknowledged.

Author information

Authors and Affiliations

  1. Department of Neurosurgery, National Institute of Mental Health and Neuro Sciences, Bangalore, 560029, India

    A. Arivazhagan & B. A. Chandramouli

  2. Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore, 560012, India

    Durairaj Mohan Kumar, Irene Rosita Pia Patric, S. Sridevi & K. Somasundaram

  3. School of Life Sciences, Jawaharlal Nehru University, New Delhi, 110067, India

    Vinay Sagar & Neelima Mondal

  4. Sri Satya Sai Institute of Higher Medical Sciences, Bangalore, 560066, India

    K. Prasanna & A. S. Hegde

  5. Biostatistics, National Institute of Mental Health and Neuro Sciences, Bangalore, 560029, India

    K. Thennarasu

  6. Neuropathology, National Institute of Mental Health and Neuro Sciences, Bangalore, 560029, India

    Balaram Thota, Mallavarapu R. Srividya & V. Santosh

  7. Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore, 560064, India

    M. R. S. Rao

  8. Molecular Reproduction, Development and Genetics, Indian Institute of Science, Bangalore, 560012, India

    P. Kondaiah

Authors
  1. A. Arivazhagan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Durairaj Mohan Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Vinay Sagar
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Irene Rosita Pia Patric
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. S. Sridevi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Balaram Thota
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Mallavarapu R. Srividya
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. K. Prasanna
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. K. Thennarasu
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Neelima Mondal
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. A. S. Hegde
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. B. A. Chandramouli
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. V. Santosh
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. M. R. S. Rao
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. P. Kondaiah
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. K. Somasundaram
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to K. Somasundaram.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOC 45 kb)

Supplementary material 2 (PDF 28 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Arivazhagan, A., Kumar, D.M., Sagar, V. et al. Higher topoisomerase 2 alpha gene transcript levels predict better prognosis in GBM patients receiving temozolomide chemotherapy: identification of temozolomide as a TOP2A inhibitor. J Neurooncol 107, 289–297 (2012). https://doi.org/10.1007/s11060-011-0758-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11060-011-0758-3

Keywords

Search

Navigation