Skip to main content
SpringerLink
Log in

HPV-type-specific response of cervical cancer cells to cisplatin after silencing replication licensing factor MCM4

  • Original Article
  • Published:
Tumor Biology

Abstract

Minichoromosome maintenance (MCM) proteins play key role in cell cycle progression by licensing DNA replication only once per cell cycle. These proteins are found to be overexpressed in cervical cancer cells. In this study, we depleted MCM4, one of the MCM 2–7 complex components by RNA interference (RNAi) in four cervical cancer cell lines. The four cell lines were selected on the basis of their human papillomavirus (HPV) infection: HPV16-positive SiHa, HPV18-positive ME-180, HPV16- and HPV18-positive CaSki, and HPV-negative C-33A. The MCM4-deficient cells irrespective of their HPV status grow for several generations and maintain regular cell cycle. We did not find any evidence of augmented response to a short-term (48 h) cisplatin treatment in these MCM4-deficient cells. However, MCM4−/HPV16+ SiHa cells cannot withstand a prolonged treatment (up to 5 days) of even a sublethal dosage of cisplatin. They show increased chromosomal instability compared to their control counterparts. On the other hand, MCM4-deficient CaSki cells (both HPV16+ and 18+) remain resistant to a prolonged exposure to cisplatin. Our study indicates that cervical cancer cells may be using excess MCMs as a backup for replicative stress; however, its regulatory mechanism is dependent on the HPV status of the cells.

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
Fig. 5

Similar content being viewed by others

References

  1. Bell SP, Dutta A. DNA replication in eukaryotic cells. Annu Rev Biochem. 2002;71:333–74.

    Article  CAS  PubMed  Google Scholar 

  2. Forsburg SL. Eukaryotic MCM, proteins: beyond replication initiation. Microbiol Mol Biol Rev. 2004;68:109–31.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Kuijjer ML, Rydbeck H, Kresse SH, Buddingh EP, Lid AB, Roelofs H, et al. Identification of osteosarcoma driver genes by integrative analysis of copy number and gene expression data. Gene Chromosomes Cancer. 2012;51:696–706.

    Article  CAS  Google Scholar 

  4. Das M, Singh S, Pradhan S, Narayan G. MCM Paradox: abundance of eukaryotic replicative helicases and genomic integrity. Mol Biol International. 2014:574850.

  5. Zhong X, Chen X, Guan X, Zhang H, Ma Y, Zhang S, et al. Overexpressions of G9a and MCM7 in esophageal squamous cell carcinoma associated with poor prognosis. Histopathology. 2015;66:192–200.

    Article  PubMed  Google Scholar 

  6. Madine MA, Swietlik M, Pelizon C, Romanowski P, Mills AD, Laskey RA. The roles of the MCM, ORC and Cdc6 proteins in determining the replication competence of chromatin in quiescent cells. J Struct Biol. 2000;129:198–210.

    Article  CAS  PubMed  Google Scholar 

  7. Blow JJ, Hodgson B. Replication licensing—defining the proliferative state? Trends Cell Biol. 2002;12:72–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Shreeram S, Sparks A, Lane DP, Blow JJ. Cell type-specific responses of human cells to inhibition of replication licensing. Oncogene. 2002;21:6624–32.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Feng D, Tu Z, Wu W, Liang C. Inhibiting the expression of DNA replication-initiation proteins induces apoptosis in human cancer cells. Cancer Res. 2003;63:7356–64.

    CAS  PubMed  Google Scholar 

  10. Liu M, Li JS, Tian DP, Huang B, Rosqvist S, Su M. MCM2 expression levels predict diagnosis and prognosis in gastric cardiac cancer. Histol Histopathol. 2013;28:481–92.

    PubMed  Google Scholar 

  11. Erkan EP, Strobel 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 

  12. Das M, Prasad SB, Yadav SS, Govardhan HB, Pandey LK, Singh S, et al. Over expression of minichromosome maintenance genes is clinically correlated to cervical carcinogenesis. PLoS One. 2013;8:e69607.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Narayan G, Arias-Pulido H, Nandula SV, Basso K, Sugirtharaj DD, Vargas H, et al. Promoter hypermethylation of FANCF: disruption of Fanconi Anemia-BRCA pathway in cervical cancer. Cancer Res. 2004;64:2994–7.

    Article  CAS  PubMed  Google Scholar 

  14. Narayan G, Bourdon V, Chaganti S, Arias-Pulido H, Nandula SV, Rao PH, et al. Gene dosage alterations revealed by cDNA microarray analysis in cervical cancer: identification of candidate amplified and overexpressed genes. Gene Chromosomes Cancer. 2007;46:373–84.

    Article  CAS  Google Scholar 

  15. Baay MF, Quint WG, Koudstaal J, Hollema H, Duk JM, Burger MP, et al. Comprehensive study of several general and type-specific primer pairs for detection of human papillomavirus DNA by PCR in paraffin-embedded cervical carcinomas. J Clin Microbiol. 1996;34:745–7.

    CAS  PubMed  PubMed Central  Google Scholar 

  16. Shukla S, Bharti AC, Mahata S, Hussain S, Hedau S, Sharma R, et al. Application of a multiplex PCR to cervical cells collected by a paper smear for the simultaneous detection of all mucosal human papillomaviruses (HPVs) and typing of high-risk HPV types 16 and 18. J Med Microbiol. 2010;59:1303–10.

    Article  CAS  PubMed  Google Scholar 

  17. Symeonidou IE, Kotsantis P, Roukos V, Rapsomaniki MA, Grecco HE, Bastiaens P, et al. Multi-step loading of human minichromosome maintenance proteins in live human cells. J Biol Chem. 2013;288:35852–67.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Woodward AM, Gohler T, Luciani MG, Oehlmann M, Ge X, Gartner A, et al. Excess Mcm2-7 license dormant origins of replication that can be used under conditions of replicative stress. J Cell Biol. 2006;173:673–83.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Alver RC, Chadha GS, Blow JJ. The contribution of dormant origins to genome stability: from cell biology to human genetics. DNA Repair (Amst). 2014;19:182–9.

    Article  CAS  Google Scholar 

  20. Ge XQ, Jackson DA, Blow JJ. Dormant origins licensed by excess Mcm2-7 are required for human cells to survive replicative stress. Genes Dev. 2007;21:3331–41.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. 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 U S A. 2008;105:8956–61.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Hausen ZH. Papillomaviruses and cancer: from basic studies to clinical application. Nat Rev Cancer. 2002;2:342–50.

    Article  PubMed  Google Scholar 

  23. Saxena A, Yashar C, Taylor DD, Gercel-Taylor C. Cellular response to chemotherapy and radiation in cervical cancer. Am J Obstet Gynecol. 2005;192:1399–403.

    Article  CAS  PubMed  Google Scholar 

  24. Nicholas S E, Schwacha A. The Mcm2-7 replicative helicase: a promising chemotherapeutic target. BioMed Res International. 2014:549719.

  25. Galluzzi L, Senovilla L, Vitale I, Michels J, Martins I, Kepp O, et al. Molecular mechanisms of cisplatin resistance. Oncogene. 2012;31:1869–83.

    Article  CAS  PubMed  Google Scholar 

  26. Vici P, Mariani L, Pizzuti L, Sergi D, Di Lauro L, Vizza E, et al. Emerging biological treatments for uterine cervical carcinoma. J Cancer. 2014;5:86–97.

    Article  PubMed  PubMed Central  Google Scholar 

  27. Tommasino M. The human papillomavirus family and its role in carcinogenesis. Sem Cancer Biol. 2014;26:13–21.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors thankfully acknowledge the financial support from the Department of Biotechnology, Government of India (vide project no. BT/PR9246/Med/30/17/2007), to GN, SS, and SP; Banaras Hindu University (XIth Plan) to SS; Council of Scientific and Industrial Research, Government of India, for research fellowship to MD; and Interdisciplinary School of Life Sciences, Faculty of Science, Banaras Hindu University for equipment facilities.

Conflicts of interest

None

Author information

Author notes

  1. Shyam Babu Prasad

    Present address: Division of Molecular Oncology, Institute of Cytology and Preventive Oncology, I-7, Sector-39, Noida, 201 301, India

Authors and Affiliations

  1. Cancer Genetics Laboratory, Department of Molecular and Human Genetics, Banaras Hindu University, Varanasi, 221 005, Uttar Pradesh, India

    Mitali Das, Shyam Babu Prasad, Suresh Singh Yadav, Arusha Modi & Gopeshwar Narayan

  2. Department of Zoology, Mahila Mahavidyalaya, Banaras Hindu University, Varanasi, 221 005, Uttar Pradesh, India

    Sunita Singh

  3. Department of Radiotherapy and Radiation Medicine, Banaras Hindu University, Varanasi, 221 005, Uttar Pradesh, India

    Satyajit Pradhan

Authors
  1. Mitali Das
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shyam Babu Prasad
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Suresh Singh Yadav
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Arusha Modi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sunita Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Satyajit Pradhan
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Gopeshwar Narayan
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Gopeshwar Narayan.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Das, M., Prasad, S.B., Yadav, S.S. et al. HPV-type-specific response of cervical cancer cells to cisplatin after silencing replication licensing factor MCM4. Tumor Biol. 36, 9987–9994 (2015). https://doi.org/10.1007/s13277-015-3782-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13277-015-3782-7

Keywords

Search

Navigation