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.

  • Article
  • Published:

MCM8 is regulated by EGFR signaling and promotes the growth of glioma stem cells through its interaction with DNA-replication-initiating factors

Oncogene volume 40, pages 4615–4624 (2021)Cite this article

Subjects

Abstract

Mini-chromosome maintenance (MCM) proteins are critical components of DNA-replication-licensing factors. MCM8 is an MCM protein that exhibits oncogenic functions in several human malignancies. However, the role of MCM8 in glioblastomas (GBMs) has remained unclear. In the present study, we investigated the biological functions and mechanisms of MCM8 in glioma stem cells (GSCs). The clinical relevance of MCM8 mRNA expression was explored via TCGA and REMBRANDT datasets. The effects of MCM8 on the self-renewal and tumorigenicity of GSCs were examined both in vitro and in vivo. The regulation of MCM8 expression and its interacting proteins were also evaluated. We found that the expression of MCM8 was elevated in high-grade gliomas and classical molecular subtypes and was inversely correlated with patient prognosis. GSCs had a significantly higher expression of MCM8 compared with that in normal glioma cells. Silencing of MCM8 induced G0/G1 arrest and apoptosis, as well as inhibited the proliferation and self-renewal of GSCs. Forced expression of MCM8 enhanced clonogenicity of GSCs both in vitro and in vivo. MCM8 expression was regulated by EGFR signaling, which was mediated by NF-κB (p65). MCM8 interacted with DNA-replication-initiating factors—including EZH2, CDC6, and CDCA2—and influenced these factors to associate with chromatin. In addition, MCM8 knockdown increased the sensitivity of GSCs to radiation and TMZ treatments. Our findings suggest that MCM8, regulated by the EGFR pathway, maintains the clonogenic and tumorigenic potential of GSCs through interaction with DNA-replication-initiating factors; hence, MCM8 may represent a novel therapeutic target in GBMs.

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

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

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

Fig. 1: Correlation between MCM8 expression and clinic-pathologic features of gliomas.
Fig. 2: MCM8 knockdown inhibits the clonogenicity of GSCs in vitro.
Fig. 3: Effect of forced MCM8 expression on in vitro GSC growth.
Fig. 4: Effect of MCM8 on in vivo GSC growth.
Fig. 5: MCM8 expression is regulated by the EGFR pathway.
Fig. 6: MCM8 interacts with EZH2, CDC6, and CDCA2.
Fig. 7: Effect of MCM8 knockdown on the therapeutic sensitivity of GSCs.
Fig. 8: A working model of MCM8-promoted cell survival and treatment resistance in glioma stem cells.

Similar content being viewed by others

References

  1. Weller M, van den Bent M, Tonn JC, Stupp R, Preusser M, Cohen-Jonathan-Moyal E, et al. European Association for Neuro-Oncology (EANO) guideline on the diagnosis and treatment of adult astrocytic and oligodendroglial gliomas. Lancet Oncol. 2017;18:e315–29.

    Article  PubMed  Google Scholar 

  2. Azzarelli R, Simons BD, Philpott A. The developmental origin of brain tumours: a cellular and molecular framework. Development. 2018;145:10.

    Article  Google Scholar 

  3. Vescovi AL, Galli R, Reynolds BA. Brain tumour stem cells. Nat Rev Cancer. 2006;6:425–36.

    Article  CAS  PubMed  Google Scholar 

  4. Freeman A, Morris LS, Mills AD, Stoeber K, Laskey RA, Williams GH, et al. Minichromosome maintenance proteins as biological markers of dysplasia and malignancy. Clin Cancer Res. 1999;5:2121–32.

    CAS  PubMed  Google Scholar 

  5. Honeycutt KA, Chen Z, Koster MI, Miers M, Nuchtern J, Hicks J, et al. Deregulated minichromosomal maintenance protein MCM7 contributes to oncogene driven tumorigenesis. Oncogene. 2006;25:4027–32.

    Article  CAS  PubMed  Google Scholar 

  6. Wang Y, Chen H, Zhang J, Cheng A, Yu J, To KF, et al. MCM family in gastrointestinal cancer and other malignancies: from functional characterization to clinical implication. Biochim Biophys Acta Rev Cancer. 2020;1874:188415.

    Article  CAS  PubMed  Google Scholar 

  7. Ishimi Y. Regulation of MCM2-7 function. Genes Genet Syst. 2018;93:125–33.

    Article  CAS  PubMed  Google Scholar 

  8. Matson JP, Dumitru R, Coryell P, Baxley RM, Chen W, Twaroski K, et al. Rapid DNA replication origin licensing protects stem cell pluripotency. Elife. 2017;6:6.

    Article  Google Scholar 

  9. He DM, Ren BG, Liu S, Tan LZ, Cieply K, Tseng G, et al. Oncogenic activity of amplified miniature chromosome maintenance 8 in human malignancies. Oncogene. 2017;36:3629–39.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Cai L, Zhao K, Yuan X. Expression of minichromosome maintenance 8 in chronic myelogenous leukemia. Int J Clin Exp Pathol. 2015;8:14180–8.

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Peng YP, Zhu Y, Yin LD, Zhang JJ, Guo S, Fu Y, et al. The expression and prognostic roles of MCMs in pancreatic cancer. PLoS One. 2016;11:e0164150.

    Article  PubMed  PubMed Central  Google Scholar 

  12. Liu Z, Li J, Chen J, Shan Q, Dai H, Xie H, et al. MCM family in HCC: MCM6 indicates adverse tumor features and poor outcomes and promotes S/G2 cell cycle progression. BMC Cancer. 2018;18:200.

    Article  PubMed  PubMed Central  Google Scholar 

  13. Erkan EP, Ströbel T, Lewandrowski G, Tannous B, Madlener S, Czech T, et al. Depletion of minichromosome maintenance protein 7 inhibits glioblastoma multiforme tumor growth in vivo. Oncogene. 2014;33:4778–85.

    Article  CAS  PubMed  Google Scholar 

  14. Hua C, Zhao G, Li Y, Bie L. Minichromosome maintenance (MCM) family as potential diagnostic and prognostic tumor markers for human gliomas. BMC Cancer. 2014;14:526.

    Article  PubMed  PubMed Central  Google Scholar 

  15. Cai HQ, Cheng ZJ, Zhang HP, Wang PF, Zhang Y, Hao JJ, et al. Overexpression of MCM6 predicts poor survival in patients with glioma. Hum Pathol. 2018;78:182–7.

    Article  CAS  PubMed  Google Scholar 

  16. 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  PubMed Central  Google Scholar 

  17. Greenall SA, McKenzie M, Seminova E, Dolezal O, Pearce L, Bentley J, et al. Most clinical anti-EGFR antibodies do not neutralize both wtEGFR and EGFRvIII activation in glioma. Neuro Oncol. 2019;21:1016–27.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Mukherjee S, Tucker-Burden C, Kaissi E, Newsam A, Duggireddy H, Chau M, et al. CDK5 inhibition resolves PKA/cAMP-independent activation of CREB1 signaling in glioma stem cells. Cell Rep. 2018;23:1651–64.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Labib K, Kearsey SE, Diffley JF. MCM2-7 proteins are essential components of prereplicative complexes that accumulate cooperatively in the nucleus during G1-phase and are required to establish, but not maintain, the S-phase checkpoint. Mol Biol Cell. 2001;12:3658–67.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Im JS, Ki SH, Farina A, Jung DS, Hurwitz J, Lee JK. Assembly of the Cdc45-Mcm2-7-GINS complex in human cells requires the Ctf4/And-1, RecQL4, and Mcm10 proteins. Proc Natl Acad Sci USA. 2009;106:15628–32.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Nishimura K, Ishiai M, Horikawa K, Fukagawa T, Takata M, Takisawa H, et al. Mcm8 and Mcm9 form a complex that functions in homologous recombination repair induced by DNA interstrand crosslinks. Mol Cell. 2012;47:511–22.

    Article  CAS  PubMed  Google Scholar 

  22. Volkening M, Hoffmann I. Involvement of human MCM8 in prereplication complex assembly by recruiting hcdc6 to chromatin. Mol Cell Biol. 2005;25:1560–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. 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;31:570–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Yang W, Xia Y, Cao Y, Zheng Y, Bu W, Zhang L, et al. EGFR-induced and PKCepsilon monoubiquitylation-dependent NF-kappaB activation upregulates PKM2 expression and promotes tumorigenesis. Mol Cell. 2012;48:771–84.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Li X, Qian X, Jiang H, Xia Y, Zheng Y, Li J, et al. Nuclear PGK1 alleviates ADP-dependent inhibition of CDC7 to promote DNA replication. Mol Cell. 2018;72:650–e8.

    Article  CAS  PubMed  Google Scholar 

  26. Liu Y, Liu S, Yuan S, Yu H, Zhang Y, Yang X, et al. Chromodomain protein CDYL is required for transmission/restoration of repressive histone marks. J Mol Cell Biol. 2017;9:178–94.

    Article  CAS  PubMed  Google Scholar 

  27. Zhang X, Xiao D, Wang Z, Zou Y, Huang L, Lin W, et al. MicroRNA-26a/b regulate DNA replication licensing, tumorigenesis, and prognosis by targeting CDC6 in lung cancer. Mol Cancer Res. 2014;12:1535–46.

    Article  CAS  PubMed  Google Scholar 

  28. Jin X, Kim LJY, Wu Q, Wallace LC, Prager BC, Sanvoranart T, et al. Targeting glioma stem cells through combined BMI1 and EZH2 inhibition. Nat Med. 2017;23:1352–61.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Friedman HS, Kerby T, Calvert H. Temozolomide and treatment of malignant glioma. Clin Cancer Res. 2000;6:2585–97.

    CAS  PubMed  Google Scholar 

  30. De Ruysscher D, Niedermann G, Burnet NG, Siva S, Lee A, Hegi-Johnson F, et al. Radiotherapy toxicity. Nat Rev Dis Prim. 2019;5:13.

    Article  PubMed  Google Scholar 

  31. Lee KY, Im JS, Shibata E, Park J, Handa N, Kowalczykowski SC, et al. MCM8-9 complex promotes resection of double-strand break ends by MRE11-RAD50-NBS1 complex. Nat Commun. 2015;6:7744.

    Article  CAS  PubMed  Google Scholar 

  32. Park J, Long DT, Lee KY, Abbas T, Shibata E, Negishi M, et al. The MCM8-MCM9 complex promotes RAD51 recruitment at DNA damage sites to facilitate homologous recombination. Mol Cell Biol. 2013;33:1632–44.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Han S, Li X, Qiu B, Jiang T, Wu A. Can lateral ventricle contact predict the ontogeny and prognosis of glioblastoma. J Neurooncol. 2015;124:45–55.

    Article  CAS  PubMed  Google Scholar 

  34. Jiang Y, Song Y, Wang R, Hu T, Zhang D, Wang Z, et al. NFAT1-mediated regulation of NDEL1 promotes growth and invasion of glioma stem-like cells. Cancer Res. 2019;79:2593–603.

    Article  CAS  PubMed  Google Scholar 

  35. Song Y, Zhang L, Jiang Y, Hu T, Zhang D, Qiao Q, et al. MTBP regulates cell survival and therapeutic sensitivity in TP53 wildtype glioblastomas. Theranostics. 2019;9:6019–30.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Han S, Wang C, Qin X, Xia J, Wu A. LPS alters the immuno-phenotype of glioma and glioma stem-like cells and induces in vivo antitumor immunity via TLR4. J Exp Clin Cancer Res. 2017;36:83.

    Article  PubMed  PubMed Central  Google Scholar 

  37. Mendez J, Stillman B. Chromatin association of human origin recognition complex, cdc6, and minichromosome maintenance proteins during the cell cycle: assembly of prereplication complexes in late mitosis. Mol Cell Biol. 2000;20:8602–12.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Wang Q, Zeng F, Sun Y, Qiu Q, Zhang J, Huang W, et al. Etk interaction with PFKFB4 modulates chemoresistance of small-cell lung cancer by regulating autophagy. Clin Cancer Res. 2018;24:950–62.

    Article  CAS  PubMed  Google Scholar 

  39. Guo S, Meng L, Liu H, Yuan L, Zhao N, Ni J, et al. Trio cooperates with Myh9 to regulate neural crest-derived craniofacial development. Theranostics. 2021;11:4316–34.

    Article  PubMed  PubMed Central  Google Scholar 

  40. Bowman RL, Wang Q, Carro A, Verhaak RG, Squatrito M. GlioVis data portal for visualization and analysis of brain tumor expression datasets. Neuro Oncol. 2017;19:139–41.

    Article  CAS  PubMed  Google Scholar 

  41. Tang Z, Li C, Kang B, Gao G, Li C, Zhang Z. GEPIA: a web server for cancer and normal gene expression profiling and interactive analyses. Nucleic Acids Res. 2017;45:W98–102.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank Qingchang Li from the Department of Pathology at China Medical University for technological support and assistance with statistical analyses. This work was supported by the LiaoNing Revitalization Talents Program (No. XLYC1807253) and the National Natural Science Foundation of China (No. 81772653 and 81402045)

Author information

Author notes

  1. These authors contributed equally: Xiaoliang Wang, Li Zhang, Yifu Song

Authors and Affiliations

  1. Department of Neurosurgery, The First Hospital of China Medical University, Shenyang, China

    Xiaoliang Wang, Li Zhang, Yifu Song, Yang Jiang, Run Wang, Tianhao Hu & Sheng Han

  2. Department of Neurosurgery, Shanghai First People’s Hospital of Shanghai Jiao Tong University School of Medicine, Shanghai, China

    Yang Jiang

  3. Department of Pathology, China Medical University, Shenyang, China

    Di Zhang

Authors
  1. Xiaoliang Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Li Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yifu Song
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yang Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Di Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Run Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Tianhao Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Sheng Han
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sheng Han.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Wang, X., Zhang, L., Song, Y. et al. MCM8 is regulated by EGFR signaling and promotes the growth of glioma stem cells through its interaction with DNA-replication-initiating factors. Oncogene 40, 4615–4624 (2021). https://doi.org/10.1038/s41388-021-01888-1

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41388-021-01888-1

This article is cited by

Search

Advanced search

Quick links