Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Short Communication
  • Published:

Replication stress and oxidative damage contribute to aberrant constitutive activation of DNA damage signalling in human gliomas

Oncogene volume 29, pages 5095–5102 (2010)Cite this article

Subjects

Abstract

Malignant gliomas, the deadliest of brain neoplasms, show rampant genetic instability and resistance to genotoxic therapies, implicating potentially aberrant DNA damage response (DDR) in glioma pathogenesis and treatment failure. Here, we report on gross, aberrant constitutive activation of DNA damage signalling in low- and high-grade human gliomas, and analyze the sources of such endogenous genotoxic stress. Based on analyses of human glioblastoma multiforme (GBM) cell lines, normal astrocytes and clinical specimens from grade II astrocytomas (n=41) and grade IV GBM (n=60), we conclude that the DDR machinery is constitutively activated in gliomas, as documented by phosphorylated histone H2AX (γH2AX), activation of the ATM-Chk2-p53 pathway, 53BP1 foci and other markers. Oxidative DNA damage (8-oxoguanine) was high in some GBM cell lines and many GBM tumors, while it was low in normal brain and grade II astrocytomas, despite the degree of DDR activation was higher in grade II tumors. Markers indicative of ongoing DNA replication stress (Chk1 activation, Rad17 phosphorylation, replication protein A foci and single-stranded DNA) were present in GBM cells under high- or low-oxygen culture conditions and in clinical specimens of both low- and high-grade tumors. The observed global checkpoint signaling, in contrast to only focal areas of overabundant p53 (indicative of p53 mutation) in grade II astrocytomas, are consistent with DDR activation being an early event in gliomagenesis, initially limiting cell proliferation (low Ki-67 index) and selecting for mutations of p53 and likely other genes that allow escape (higher Ki-67 index) from the checkpoint and facilitate tumor progression. Overall, these results support the potential role of the DDR machinery as a barrier to gliomagenesis and indicate that replication stress, rather than oxidative stress, fuels the DNA damage signalling in early stages of astrocytoma development.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4

Similar content being viewed by others

References

  • Bao S, Wu Q, McLendon RE, Hao Y, Shi Q, Hjelmeland AB et al. (2006). Glioma stem cells promote radio resistance by preferential activation of the DNA damage response. Nature 444: 756–760.

    Article  CAS  PubMed  Google Scholar 

  • Bartek J, Bartkova J, Lukas J . (2007). DNA damage signaling guards against activated oncogenes and tumour progression. Oncogene 26: 7773–7779.

    Article  CAS  PubMed  Google Scholar 

  • Bartkova J, Bakkenist CJ, Rajpert-De Meyts E, Skakkebaek NE, Sehested M, Lukas J et al. (2005a). ATM activation in normal human tissues and testicular cancer. Cell Cycle 4: 838–845.

    Article  CAS  PubMed  Google Scholar 

  • Bartkova J, Horejsi Z, Koed K, Krämer A, Tort F, Zieger K et al. (2005b). DNA damage response as a candidate anti-cancer barrier in early human tumorigenesis. Nature 434: 864–870.

    Article  CAS  PubMed  Google Scholar 

  • Bartkova J, Horejsi Z, Sehested M, Nesland JM, Rajpert-De Meyts E, Skakebaek NE et al. (2007). DNA damage response mediators MDC1 and 53 BP1: constitutive activation and aberrant loss in breast and lung cancer, but not in testicular germ cell tumours. Oncogene 26: 7414–7422.

    Article  CAS  PubMed  Google Scholar 

  • Bartkova J, Rezaei N, Liontos M, Karakaidos P, Kletsas D, Issaeva LV et al. (2006). Oncogene-induced senescence is part of the tumorigenesis barrier imposed by DNA damage checkpoints. Nature 444: 633–637.

    Article  CAS  PubMed  Google Scholar 

  • Bekker-Jensen S, Lukas C, Melander F, Bartek J, Lukas J . (2005). Dynamic assembly and sustained retention of 53BP1 at the sites of DNA damage are controlled by Mdc1/NFBD1. J Cell Biol 170: 201–211.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bonner WM, Redon CE, Dickey JS, Nakamura AJ, Sedelnikova OA, Solier S et al. (2008). H2AX and cancer. Nat Rev Cancer 8: 957–967.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bristow RG, Hill RP . (2008). Hypoxia and metabolism. Hypoxia, DNA repair and genetic instability. Nat Rev Cancer 8: 180–192.

    Article  CAS  PubMed  Google Scholar 

  • Cimprich KA, Cortez D . (2008). ATR: An essential regulator of genome integrity. Nat Rev Mol Cell Biol 9: 616–627.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Di Micco R, Fumagalli M, Cicalese A, Piccinin S, Gasparini P, Luise C et al. (2006). Oncogene-induced senescence is a DNA damage response triggered by DNA hyper-replication. Nature 444: 638–642.

    Article  CAS  PubMed  Google Scholar 

  • DiTullio RA, Mochan TA, Venere M, Bartkova J, Sehested M, Bartek J et al. (2002). 53BP1 functions in an ATM-dependent checkpoint pathway that is constitutively activated in human cancer. Nat Cell Biol 4: 998–1002.

    Article  CAS  PubMed  Google Scholar 

  • Dungey FA, Caldecott KW, Chalmers AJ . (2009). Enhanced radiosensitization of human glioma cells by combining inhibition of poly(ADP-ribose) polymerase with inhibition of heat shock protein 90. Mol Cancer Ther 8: 2243–2254.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Furnari FB, Fenton T, Bachoo RM, Mukasa A, Stommel JM, Stegh A et al. (2007). Malignant astrocytic glioma; genetics, biology and paths to treatment. Genes & Dev 21: 2683–2710.

    Article  CAS  Google Scholar 

  • Gorgoulis VG, Vassiliou LV, Karakaidos P, Zacharatos P, Kotsinas A, Liliglou T et al. (2005). Activation of the DNA damage checkpoint and genomic instability in human precancerous lesions. Nature 434: 907–913.

    Article  CAS  PubMed  Google Scholar 

  • Halazonetis TD, Gorgoulis VG, Bartek J . (2008). An oncogene-induced DNA damage model for cancer development. Science 319: 1352–1355.

    Article  CAS  PubMed  Google Scholar 

  • Helleday T, Petermann E, Lundin C, Hodgson B, Sharma RA . (2008). DNA repair pathways as targets for cancer therapy. Nat Rev Cancer 8: 193–204.

    Article  CAS  PubMed  Google Scholar 

  • Jackson SP, Bartek J . (2009). The DNA-damage response in human biology and disease. Nature 461: 1071–1078.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Knizetova P, Ehrmann J, Hlobilkova A, Vancova I, Kalita O, Kolar Z et al. (2008). Autocrine regulation of glioblastoma cell cycle progression, viability and radiorestance through the VEGF-VEGFR2 (KDR) interplay. Cell Cycle 7: 2553–2561.

    Article  CAS  PubMed  Google Scholar 

  • Li D, Marks JD, Schumacker PT, Young RM, Brorson JR . (2005). Physiological hypoxia promotes survival of cultured cortical neurons. Eur J Neurosci 6: 1319–1326.

    Article  Google Scholar 

  • Luo J, Solimini NL, Elledge SJ . (2009). Principles of cancer therapy: oncogene and non-oncogene addiction. Cell 6: 823–837.

    Article  Google Scholar 

  • Malette FA, Ferbeyre G . (2007). The DNA damage signaling pathway connects oncogenis stress to cellular senescence. Cell Cycle 6: 1831–1836.

    Article  Google Scholar 

  • Martin SA, Lord CJ, Ashworth A . (2008). DNA repair deficiency as a terapeutic target in cancer. Curr Opin Genet Dev 18: 80–86.

    Article  CAS  PubMed  Google Scholar 

  • Mistrik M, Oplustilova L, Lukas J, Bartek J . (2009). Low-dose DNA damage and replications stress responses quantified by optimized automated single-cell image analysis. Cell Cycle 8: 2592–2599.

    Article  CAS  PubMed  Google Scholar 

  • Mukherjee B, McEllin B, Camacho CV, Tomimatsu N, Sirasanagandala S, Nannepaga S et al. (2009). EGFRvIII and DNA double-strand break repair: A molecular mechanism for radioresistance in glioblastoma. Cancer Res 69: 4252–4259.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nuciforo PG, Luise C, Capra M, Pelosi G, d'Adda di Fagagna F . (2007). Complex engagement of DNA-damage response pathways in human cancer and in lung tumor progression. Carcinogenesis 28: 2082–2088.

    Article  CAS  PubMed  Google Scholar 

  • Raderschall E, Golub EI, Haaf T . (1999). Nuclear foci of mammalian recombination proteins are located at single-stranded DNA regions formed after DNA damage. Pro Natl Acad Sci USA 96: 1921–1926.

    Article  CAS  Google Scholar 

  • The Cancer Genome Atlas Research Network (2008). Comprehensive genomic characterization defines human glioblastoma genes and core pathways. Nature 455: 1061–1068.

    Article  Google Scholar 

  • Zheng H, Ying H, Yan H, Kimmelman AC, Hiller DJ, Chen AJ et al. (2008). P53 and Pten control neural and glioma stem/progenitor cell renewal and differentiation. Nature 455: 1129–1134.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhou B-BS, Bartek J . (2004). Targetting the checkpoint kinases. Chemosensitization versus chemoprotection. Nat Rev Cancer 4: 216–225.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank J Darling, University of Wolverhampton and CV Bree, University of Amsterdam for donating the SS859 and Gli-6 cells, respectively and MyungHee Lee for excellent technical assistance. Grant support was obtained from Danish Cancer Society, Danish National Research Foundation, Danish Research Council, Vilhelm Pedersen and Hustrus Mindelegat, Czech Ministry of Education (MSMT6198959216), Grant Agency of the Czech Ministry of Health (NS10282-3/2009), Lundbeck Foundation (R13-A1287) and European Commission (projects: CZ.1.05/2.1.00/01.0030, Active p53, Infla-Care, GENICA).

Author information

Author notes

  1. J Bartkova and P Hamerlik: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute of Cancer Biology and Centre for Genotoxic Stress Research, Danish Cancer Society, Copenhagen, Denmark

    J Bartkova, P Hamerlik, J Lukas & J Bartek

  2. Laboratory of Genome Integrity, Institute of Molecular and Translational Medicine, Palacky University, Olomouc, Czech Republic

    P Hamerlik & J Bartek

  3. Molecular Pathology, Institute of Molecular and Translational Medicine, Palacky University, Olomouc, Czech Republic

    P Hamerlik, J Ehrmann, A Hlobilkova & Z Kolar

  4. Department of Radiation Biology, Copenhagen University Hospital, Copenhagen, Denmark

    M-T Stockhausen & H S Poulsen

  5. Department of Neuropathology, Copenhagen University Hospital, Copenhagen, Denmark

    H Laursen & H Broholm

  6. Department of Neurosurgery, University Hospital, Olomouc, Czech Republic

    O Kalita

Authors
  1. J Bartkova
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. P Hamerlik
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M-T Stockhausen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. J Ehrmann
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. A Hlobilkova
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. H Laursen
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. O Kalita
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Z Kolar
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. H S Poulsen
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. H Broholm
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. J Lukas
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. J Bartek
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to J Lukas or J Bartek.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies the paper on the Oncogene website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bartkova, J., Hamerlik, P., Stockhausen, MT. et al. Replication stress and oxidative damage contribute to aberrant constitutive activation of DNA damage signalling in human gliomas. Oncogene 29, 5095–5102 (2010). https://doi.org/10.1038/onc.2010.249

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2010.249

Keywords

This article is cited by

Search

Advanced search

Quick links