Skip to main content
SpringerLink
Log in

MiR-338-5p sensitizes glioblastoma cells to radiation through regulation of genes involved in DNA damage response

Tumor Biology volume 37, pages 7719–7727 (2016)Cite this article

Abstract

Glioblastoma multiforme (GBM) is the most aggressive form of brain tumor. Despite radical surgery and radiotherapy supported by chemotherapy, the disease still remains incurable with an extremely low median survival rate of 12–15 months from the time of initial diagnosis. The main cause of treatment failure is considered to be the presence of cells that are resistant to the treatment. MicroRNAs (miRNAs) as regulators of gene expression are involved in the tumor pathogenesis, including GBM. MiR-338 is a brain-specific miRNA which has been described to target pathways involved in proliferation and differentiation. In our study, miR-338-3p and miR-338-5p were differentially expressed in GBM tissue in comparison to non-tumor brain tissue. Overexpression of miR-338-3p with miRNA mimic did not show any changes in proliferation rates in GBM cell lines (A172, T98G, U87MG). On the other hand, pre-miR-338-5p notably decreased proliferation and caused cell cycle arrest. Since radiation is currently the main treatment modality in GBM, we combined overexpression of pre-miR-338-5p with radiation, which led to significantly decreased cell proliferation, increased cell cycle arrest, and apoptosis in comparison to irradiation-only cells. To better elucidate the mechanism of action, we performed gene expression profiling analysis that revealed targets of miR-338-5p being Ndfip1, Rheb, and ppp2R5a. These genes have been described to be involved in DNA damage response, proliferation, and cell cycle regulation. To our knowledge, this is the first study to describe the role of miR-338-5p in GBM and its potential to improve the sensitivity of GBM to radiation.

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

Access options

Buy single article

Instant access to the full article PDF.

USD 39.95

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

References

  1. Louis DN, Ohgaki H, Wiestler OD, Cavenee WK, Burger PC, Jouvet A, et al. The 2007 WHO classification of tumours of the central nervous system. Acta Neuropathol. 2007;114:97–109.

    Article  PubMed  PubMed Central  Google Scholar 

  2. Stupp R, Mason WP, van den Bent MJ, Weller M, Fisher B, Taphoorn MJ, et al. Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med. 2005;352:987–96.

    Article  CAS  PubMed  Google Scholar 

  3. Ahmed SU, Carruthers R, Gilmour L, Yildirim S, Watts C, Chalmers AJ. Selective inhibition of parallel DNA damage response pathways optimizes radiosensitization of glioblastoma stem-like cells. Cancer Res. 2015;75:4416–28.

    Article  CAS  PubMed  Google Scholar 

  4. Niyazi M, Siefert A, Schwarz SB, Ganswindt U, Kreth FW, Tonn JC, et al. Therapeutic options for recurrent malignant glioma. Radiother Oncol. 2011;98:1–14.

    Article  PubMed  Google Scholar 

  5. Huang Q, Zhang QB, Dong J, Wu YY, Shen YT, Zhao YD, et al. Glioma stem cells are more aggressive in recurrent tumors with malignant progression than in the primary tumor, and both can be maintained long-term in vitro. BMC Cancer. 2008;8:304.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Walbert T, Mikkelsen T. Recurrent high-grade glioma: a diagnostic and therapeutic challenge. Expert Rev Neurother. 2011;11:509–18.

    Article  PubMed  Google Scholar 

  7. Squatrito M, Holland EC. DNA damage response and growth factor signaling pathways in gliomagenesis and therapeutic resistance. Cancer Res. 2011;71:5945–9.

    Article  CAS  PubMed  Google Scholar 

  8. Mao H, Lebrun DG, Yang J, Zhu VF, Li M. Deregulated signaling pathways in glioblastoma multiforme: molecular mechanisms and therapeutic targets. Cancer Investig. 2012;30:48–56.

    Article  Google Scholar 

  9. Li WB, Ma MW, Dong LJ, Wang F, Chen LX, Li XR. Microrna-34a targets notch1 and inhibits cell proliferation in glioblastoma multiforme. Cancer Biol Ther. 2011;12:477–83.

    Article  CAS  PubMed  Google Scholar 

  10. Lim YC, Roberts TL, Day BW, Stringer BW, Kozlov S, Fazry S, Bruce ZC, Ensbey KS, Walker DG, Boyd AW, Lavin MF. Increased sensitivity to ionizing radiation by targeting the homologous recombination pathway in glioma initiating cells. Mol Oncol. 2014

  11. Eramo A, Ricci-Vitiani L, Zeuner A, Pallini R, Lotti F, Sette G, et al. Chemotherapy resistance of glioblastoma stem cells. Cell Death Differ. 2006;13:1238–41.

    Article  CAS  PubMed  Google Scholar 

  12. Koshkin PA, Chistiakov DA, Chekhonin VP. Role of micrornas in mechanisms of glioblastoma resistance to radio- and chemotherapy. Biochem Biokhim. 2013;78:325–34.

    Article  CAS  Google Scholar 

  13. Besse A, Sana J, Fadrus P, Slaby O. Micrornas involved in chemo- and radioresistance of high-grade gliomas. Tumour Biol J Int Soc Oncodevelop Biol Med. 2013;34:1969–78.

    Article  CAS  Google Scholar 

  14. Croce CM. Causes and consequences of microRNA dysregulation in cancer. Nat Rev Genet. 2009;10:704–14.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Aschrafi A, Schwechter AD, Mameza MG, Natera-Naranjo O, Gioio AE, Kaplan BB. Microrna-338 regulates local cytochrome c oxidase IV mRNA levels and oxidative phosphorylation in the axons of sympathetic neurons. J Neurosci Off J Soc Neurosci. 2008;28:12581–90.

    Article  CAS  Google Scholar 

  16. Chen X, Pan M, Han L, Lu H, Hao X, Dong Q. Mir-338-3p suppresses neuroblastoma proliferation, invasion and migration through targeting prex2a. FEBS Lett. 2013;587:3729–37.

    Article  CAS  PubMed  Google Scholar 

  17. Liu C, Wang Z, Wang Y, Gu W. Mir-338 suppresses the growth and metastasis of OSCC cells by targeting nrp1. Mol Cell Biochem. 2014

  18. Sana J, Radova L, Lakomy R, Kren L, Fadrus P, Smrcka M, et al. Risk score based on microRNA expression signature is independent prognostic classifier of glioblastoma patients. Carcinogenesis. 2014;35:2756–62.

    Article  CAS  PubMed  Google Scholar 

  19. Edgar R, Domrachev M, Lash AE. Gene Expression Omnibus: NCBI gene expression and hybridization array data repository. Nucleic Acids Res. 2002;30:207–10.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Smyth GK. Limma: linear models for microarray data. In: Gentleman R, Carey VJ, Huber W, Irizarry RA, Dudoit S, editors. Bioinformatics and computational biology solutions using r and bioconductor. New York: Springer; 2005. p. 397–420.

    Chapter  Google Scholar 

  21. Otomo T, Hishii M, Arai H, Sato K, Sasai K. Microarray analysis of temporal gene responses to ionizing radiation in two glioblastoma cell lines: up-regulation of DNA repair genes. J Radiat Res. 2004;45:53–60.

    Article  CAS  PubMed  Google Scholar 

  22. Ragusa M, Majorana A, Banelli B, Barbagallo D, Statello L, Casciano I, et al. Mir152, mir200b, and miR-338, human positional and functional neuroblastoma candidates, are involved in neuroblast differentiation and apoptosis. J Mol Med (Berl). 2010;88:1041–53.

    Article  CAS  Google Scholar 

  23. Manning BD, Cantley LC. Rheb fills a GAP between TSC and TOR. Trends Biochem Sci. 28:573–6.

  24. Li X, Nan A, Xiao Y, Chen Y, Lai Y. PP2A-B56 complex is involved in dephosphorylation of gamma-H2AX in the repair process of CPT-induced DNA double-strand breaks. Toxicology. 2015;331:57–65.

    Article  CAS  PubMed  Google Scholar 

  25. Low LH, Chow YL, Li Y, Goh CP, Putz U, Silke J, et al. Nedd4 family interacting protein 1 (Ndfip1) is required for ubiquitination and nuclear trafficking of BRCA1-associated ATM activator 1 (BRAT1) during the DNA damage response. J Biol Chem. 2015;290:7141–50.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Li Y, Low LH, Putz U, Goh CP, Tan SS, Howitt J. Rab5 and Ndfip1 are involved in Pten ubiquitination and nuclear trafficking. Traffic. 2014;15:749–61.

    Article  CAS  PubMed  Google Scholar 

  27. Howitt J, Lackovic J, Low LH, Naguib A, Macintyre A, Goh CP, et al. Ndfip1 regulates nuclear Pten import in vivo to promote neuronal survival following cerebral ischemia. J Cell Biol. 2012;196:29–36.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Kalev P, Simicek M, Vazquez I, Munck S, Chen L, Soin T, et al. Loss of PPP2R2A inhibits homologous recombination DNA repair and predicts tumor sensitivity to PARP inhibition. Cancer Res. 2012;72:6414–24.

    Article  CAS  PubMed  Google Scholar 

  29. Shrivastav M, De Haro LP, Nickoloff JA. Regulation of DNA double-strand break repair pathway choice. Cell Res. 2008;18:134–47.

    Article  CAS  PubMed  Google Scholar 

  30. Rogakou EP, Pilch DR, Orr AH, Ivanova VS, Bonner WM. DNA double-stranded breaks induce histone H2AX phosphorylation on serine 139. J Biol Chem. 1998;273:5858–68.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by grants NT13514-4/2012 and NT13581-4/2012 of the Czech Ministry of Health, the project “CEITEC–Central European Institute of Technology” (CZ.1.05/1.1.00/02.0068), and the project MZ CR–RVO (MOU, 00209805).

Author information

Author notes

    Authors and Affiliations

    1. Masaryk Memorial Cancer Institute, Department of Comprehensive Cancer Care, Faculty of Medicine, Masaryk University, Brno, Czech Republic

      Andrej Besse, Jiri Sana, Radek Lakomy, Marek Svoboda & Ondrej Slaby

    2. Central European Institute of Technology (CEITEC), Masaryk University, University Campus Bohunice, Building A3, Kamenice 5, 625 00, Brno, Czech Republic

      Andrej Besse, Jiri Sana & Ondrej Slaby

    3. University Hospital Brno, Department of Pathology, Faculty of Medicine, Masaryk University, Brno, Czech Republic

      Leos Kren

    4. University Hospital Brno, Department of Neurosurgery, Faculty of Medicine, Masaryk University, Brno, Czech Republic

      Pavel Fadrus & Martin Smrcka

    5. First Department of Pathological Anatomy, St. Anne’s University Hospital and Faculty of Medicine, Masaryk University, Brno, Czech Republic

      Marketa Hermanova

    6. Department of Neurosurgery, St. Anne’s University Hospital and Faculty of Medicine, Masaryk University, Brno, Czech Republic

      Radim Jancalek

    7. Department of Neurosurgery, University Hospital Ostrava, Ostrava, Czech Republic

      Stefan Reguli & Radim Lipina

    8. Department of Radiation Oncology, Memorial Cancer Institute, Brno, Czech Republic

      Pavel Slampa

    Authors
    1. Andrej Besse
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Jiri Sana
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. Radek Lakomy
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. Leos Kren
      View author publications

      You can also search for this author in PubMed Google Scholar

    5. Pavel Fadrus
      View author publications

      You can also search for this author in PubMed Google Scholar

    6. Martin Smrcka
      View author publications

      You can also search for this author in PubMed Google Scholar

    7. Marketa Hermanova
      View author publications

      You can also search for this author in PubMed Google Scholar

    8. Radim Jancalek
      View author publications

      You can also search for this author in PubMed Google Scholar

    9. Stefan Reguli
      View author publications

      You can also search for this author in PubMed Google Scholar

    10. Radim Lipina
      View author publications

      You can also search for this author in PubMed Google Scholar

    11. Marek Svoboda
      View author publications

      You can also search for this author in PubMed Google Scholar

    12. Pavel Slampa
      View author publications

      You can also search for this author in PubMed Google Scholar

    13. Ondrej Slaby
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding author

    Correspondence to Ondrej Slaby.

    Ethics declarations

    Informed consent was obtained from each patient before the treatment.

    Conflicts of interest

    None

    Additional information

    Andrej Besse and Jiri Sana contributed equally to this work.

    Electronic supplementary material

    Below is the link to the electronic supplementary material.

    ESM 1

    (DOCX 179 kb)

    Rights and permissions

    Reprints and Permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Besse, A., Sana, J., Lakomy, R. et al. MiR-338-5p sensitizes glioblastoma cells to radiation through regulation of genes involved in DNA damage response. Tumor Biol. 37, 7719–7727 (2016). https://doi.org/10.1007/s13277-015-4654-x

    Download citation

    • Received:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s13277-015-4654-x

    Keywords

    search

    Navigation