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.

  • Original Article
  • Published:

Depletion of minichromosome maintenance protein 7 inhibits glioblastoma multiforme tumor growth in vivo

Oncogene volume 33, pages 4778–4785 (2014)Cite this article

Subjects

Abstract

Minichromosome maintenance (MCM) proteins are key elements that function as a part of the pre-replication complex to initiate DNA replication in eukaryotes. Consistent with their roles in initiating DNA replication, overexpression of MCM family members has been observed in several malignancies. Through bioinformatic analysis of The Cancer Genome Atlas’s data on glioblastoma multiforme (GBM), we found that the genomic region containing MCM7 gene was amplified in more than 80% of the present cases. To validate this finding and to identify the possible contribution of the remaining members of the MCM family to GBM progression, we used quantitative real-time PCR to analyze the gene expression profiles of all MCM family members in Grade IV (GBM) tissue samples and observed a significant upregulation in GBM samples compared with normal white matter tissues. In addition, we compared the observed gene expression profiles with those of Grade II and Grade III astrocytoma samples and determined that the observed upregulation was restricted and specific to Grade IV. MCM7 was the most upregulated gene in the gene set we analyzed, and therefore we wanted to identify the role of MCM7 in GBM progression. We determined that siRNA-mediated knockdown of MCM7 expression reduced GBM cell proliferation and also inhibited tumor growth in both xenograft and orthotopic mouse models of GBM. Taken together, our data suggest that MCM7 can be a potential prognostic marker and a novel therapeutic target in GBM therapy.

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
Figure 5
Figure 6
Figure 7

Similar content being viewed by others

References

  1. Bell SP, Stillman B . ATP-dependent recognition of eukaryotic origins of DNA replication by a multiprotein complex. Nature 1992; 357: 128–134.

    Article  CAS  Google Scholar 

  2. Donovan S, Harwood J, Drury LS, Diffley JF . Cdc6p-dependent loading of Mcm proteins onto pre-replicative chromatin in budding yeast. Proc Natl Acad Sci USA 1997; 94: 5611–5616.

    Article  CAS  Google Scholar 

  3. Randell JC, Bowers JL, Rodriguez HK, Bell SP . Sequential ATP hydrolysis by Cdc6 and ORC directs loading of the Mcm2-7 helicase. Mol Cell 2006; 21: 29–39.

    Article  CAS  Google Scholar 

  4. Maine GT, Sinha P, Tye BK . Mutants of S. cerevisiae defective in the maintenance of minichromosomes. Genetics 1984; 106: 365–385.

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Gozuacik D, Chami M, Lagorce D, Faivre J, Murakami Y, Poch O et al. Identification and functional characterization of a new member of the human Mcm protein family: hMcm8. Nucleic Acids Res 2003; 31: 570–579.

    Article  CAS  PubMed Central  Google Scholar 

  6. Maiorano D, Cuvier O, Danis E, Mechali M . MCM8 is an MCM2-7-related protein that functions as a DNA helicase during replication elongation and not initiation. Cell 2005; 120: 315–328.

    Article  CAS  Google Scholar 

  7. Lutzmann M, Maiorano D, Mechali M . Identification of full genes and proteins of MCM9, a novel, vertebrate-specific member of the MCM2-8 protein family. Gene 2005; 362: 51–56.

    Article  CAS  Google Scholar 

  8. Yoshida K . Identification of a novel cell-cycle-induced MCM family protein MCM9. Biochem Biophys Res Commun 2005; 331: 669–674.

    Article  CAS  Google Scholar 

  9. Forsburg SL . Eukaryotic MCM proteins: beyond replication initiation. Microbiol Mol Bio Rev 2004; 68: 109–131.

    Article  CAS  Google Scholar 

  10. Maiorano D, Lutzmann M, Mechali M . MCM proteins and DNA replication. Curr Opin Cell Biol 2006; 18: 130–136.

    Article  CAS  Google Scholar 

  11. Saydam O, Senol O, Schaaij-Visser TB, Pham TV, Piersma SR, Stemmer-Rachamimov AO et al. Comparative protein profiling reveals minichromosome maintenance (MCM) proteins as novel potential tumor markers for meningiomas. J Proteome Res 2010; 9: 485–494.

    Article  CAS  PubMed Central  Google Scholar 

  12. Lau KM, Chan QK, Pang JC, Li KK, Yeung WW, Chung NY et al. Minichromosome maintenance proteins 2, 3 and 7 in medulloblastoma: overexpression and involvement in regulation of cell migration and invasion. Oncogene 2010; 29: 5475–5489.

    Article  CAS  Google Scholar 

  13. Gruber-Olipitz M, Yang JW, Stroebel T, Slavc I, Lubec G . The medulloblastoma cell line DAOY but not eleven other tumor cell lines expresses minichromosome maintenance protein 4. Cancer lett 2006; 238: 76–84.

    Article  CAS  Google Scholar 

  14. Toyokawa G, Masuda K, Daigo Y, Cho HS, Yoshimatsu M, Takawa M et al. Minichromosome Maintenance Protein 7 is a potential therapeutic target in human cancer and a novel prognostic marker of non-small cell lung cancer. Mol Cancer 2011; 10: 65.

    Article  CAS  PubMed Central  Google Scholar 

  15. Kikuchi J, Kinoshita I, Shimizu Y, Kikuchi E, Takeda K, Aburatani H et al. Minichromosome maintenance (MCM) protein 4 as a marker for proliferation and its clinical and clinicopathological significance in non-small cell lung cancer. Lung Cancer 2011; 72: 229–237.

    Article  Google Scholar 

  16. Yang J, Ramnath N, Moysich KB, Asch HL, Swede H, Alrawi SJ et al. Prognostic significance of MCM2 Ki-67 and gelsolin in non-small cell lung cancer. BMC cancer 2006; 6: 203.

    Article  PubMed Central  Google Scholar 

  17. Ramnath N, Hernandez FJ, Tan DF, Huberman JA, Natarajan N, Beck AF et al. MCM2 is an independent predictor of survival in patients with non-small-cell lung cancer. J Clin Oncol 2001; 19: 4259–4266.

    Article  CAS  Google Scholar 

  18. Nishihara K, Shomori K, Fujioka S, Tokuyasu N, Inaba A, Osaki M et al. Minichromosome maintenance protein 7 in colorectal cancer: implication of prognostic significance. Int J Oncol 2008; 33: 245–251.

    CAS  PubMed  Google Scholar 

  19. Giaginis C, Giagini A, Tsourouflis G, Gatzidou E, Agapitos E, Kouraklis G et al. MCM-2 and MCM-5 expression in gastric adenocarcinoma: clinical significance and comparison with Ki-67 proliferative marker. Dig Dis Sci 2011; 56: 777–785.

    Article  CAS  Google Scholar 

  20. Tamura T, Shomori K, Haruki T, Nosaka K, Hamamoto Y, Shiomi T et al. Minichromosome maintenance-7 and geminin are reliable prognostic markers in patients with oral squamous cell carcinoma: immunohistochemical study. J Oral Pathol Med 2010; 39: 328–334.

    CAS  PubMed  Google Scholar 

  21. Guida T, Salvatore G, Faviana P, Giannini R, Garcia-Rostan G, Provitera L et al. Mitogenic effects of the up-regulation of minichromosome maintenance proteins in anaplastic thyroid carcinoma. J Clin Endocrinol Metab 2005; 90: 4703–4709.

    Article  CAS  Google Scholar 

  22. Giaginis C, Vgenopoulou S, Vielh P, Theocharis S . MCM proteins as diagnostic and prognostic tumor markers in the clinical setting. Histol Histopathol 2010; 25: 351–370.

    CAS  PubMed  Google Scholar 

  23. Louis DN, Ohgaki H, Wiestler OD, Cavenee WK . WHO Classification of Tumours of the Central Nervous System Fourth edn IARC, 2007. 312 p).

    Google Scholar 

  24. Verhaak RG, Hoadley KA, Purdom E, Wang V, Qi Y, Wilkerson MD et al. Integrated genomic analysis identifies clinically relevant subtypes of glioblastoma characterized by abnormalities in PDGFRA, IDH1, EGFR, and NF1. Cancer Cell 2010; 17: 98–110.

    Article  CAS  PubMed Central  Google Scholar 

  25. Research TCGA, Comprehensive genomic characterization defines human glioblastoma genes and core pathways. Nature 2008; 455: 1061–1068.

    Article  Google Scholar 

  26. Facoetti A, Ranza E, Benericetti E, Ceroni M, Tedeschi F, Nano R . Minichromosome maintenance protein 7: a reliable tool for glioblastoma proliferation index. Anticancer Res 2006; 26: 1071–1075.

    CAS  PubMed  Google Scholar 

  27. Facoetti A, Ranza E, Grecchi I, Benericetti E, Ceroni M, Morbini P et al. Immunohistochemical evaluation of minichromosome maintenance protein 7 in astrocytoma grading. Anticancer Res 2006; 26: 3513–3516.

    CAS  PubMed  Google Scholar 

  28. Ibarra A, Schwob E, Mendez J . Excess MCM proteins protect human cells from replicative stress by licensing backup origins of replication. Proc Natl Acad Sci USA 2008; 105: 8956–8961.

    Article  CAS  Google Scholar 

  29. Gambus A, Jones RC, Sanchez-Diaz A, Kanemaki M, van Deursen F, Edmondson RD et al. GINS maintains association of Cdc45 with MCM in replisome progression complexes at eukaryotic DNA replication forks. Nat Cell Biol 2006; 8: 358–366.

    Article  CAS  Google Scholar 

  30. Bruck I, Kaplan D . Dbf4-Cdc7 phosphorylation of Mcm2 is required for cell growth. J Biol Chem 2009; 284: 28823–28831.

    Article  CAS  PubMed Central  Google Scholar 

  31. Montagnoli A, Valsasina B, Brotherton D, Troiani S, Rainoldi S, Tenca P et al. Identification of Mcm2 phosphorylation sites by S-phase-regulating kinases. J Biol Chem 2006; 281: 10281–10290.

    Article  CAS  Google Scholar 

  32. Chuang LC, Teixeira LK, Wohlschlegel JA, Henze M, Yates JR, Mendez J et al. Phosphorylation of Mcm2 by Cdc7 promotes pre-replication complex assembly during cell-cycle re-entry. Mol Cell 2009; 35: 206–216.

    Article  CAS  PubMed Central  Google Scholar 

  33. Tsuji T, Ficarro SB, Jiang W . Essential role of phosphorylation of MCM2 by Cdc7/Dbf4 in the initiation of DNA replication in mammalian cells. Mol Biol Cell 2006; 17: 4459–4472.

    Article  CAS  PubMed Central  Google Scholar 

  34. Cho WH, Lee YJ, Kong SI, Hurwitz J, Lee JK . CDC7 kinase phosphorylates serine residues adjacent to acidic amino acids in the minichromosome maintenance 2 protein. Proc Natl Acad Sci USA 2006; 103: 11521–11526.

    Article  CAS  Google Scholar 

  35. Montagnoli A, Moll J, Colotta F . Targeting cell division cycle 7 kinase: a new approach for cancer therapy. Clin Cancer Res 2010; 16: 4503–4508.

    Article  CAS  Google Scholar 

  36. Saydam O, Senol O, Wurdinger T, Mizrak A, Ozdener GB, Stemmer-Rachamimov AO et al. miRNA-7 attenuation in Schwannoma tumors stimulates growth by upregulating three oncogenic signaling pathways. Cancer Res 2011; 71: 852–861.

    Article  CAS  Google Scholar 

  37. Saydam O, Shen Y, Wurdinger T, Senol O, Boke E, James MF et al. Downregulated microRNA-200a in meningiomas promotes tumor growth by reducing E-cadherin and activating the Wnt/beta-catenin signaling pathway. Mol Cell Biol 2009; 29: 5923–5940.

    Article  CAS  PubMed Central  Google Scholar 

  38. Saydam O, Glauser DL, Heid I, Turkeri G, Hilbe M, Jacobs AH et al. Herpes simplex virus 1 amplicon vector-mediated siRNA targeting epidermal growth factor receptor inhibits growth of human glioma cells in vivo. Mole Ther: 2005; 12: 803–812.

    Article  CAS  Google Scholar 

  39. Wurdinger T, Badr C, Pike L, de Kleine R, Weissleder R, Breakefield XO et al. A secreted luciferase for ex vivo monitoring of in vivo processes. Nat Methods 2008; 5: 171–173.

    Article  CAS  PubMed Central  Google Scholar 

Download references

Acknowledgements

This work was supported , in part, by Forschungsgesellschaft for Brain Tumors (OS, NS), EU-FP7-PEOPLE-2011-CIG (OS) and Association for Conduct of Scientific Research in the Field of Neonatology and Pediatric Intensive Care: ‘Unser Kind’ (OS and NS).

Author information

Author notes

  1. E P Erkan and T Ströbel: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Pediatrics, Molecular Neuro-Oncology Research Unit, Medical University of Vienna, Vienna, Austria

    E P Erkan, S Madlener, N Saydam & O Saydam

  2. Institute of Neurology, Medical University of Vienna, Vienna, Austria

    T Ströbel

  3. Experimental Therapeutics and Molecular Imaging Laboratory, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA

    G Lewandrowski & B Tannous

  4. Department of Neurosurgery, Medical University of Vienna, Vienna, Austria

    T Czech

Authors
  1. E P Erkan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. T Ströbel
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. G Lewandrowski
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. B Tannous
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. S Madlener
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. T Czech
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. N Saydam
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. O Saydam
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to O Saydam.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies this paper on the Oncogene website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Erkan, E., Ströbel, T., Lewandrowski, G. et al. Depletion of minichromosome maintenance protein 7 inhibits glioblastoma multiforme tumor growth in vivo. Oncogene 33, 4778–4785 (2014). https://doi.org/10.1038/onc.2013.423

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2013.423

Keywords

This article is cited by

Search

Advanced search

Quick links