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Role of miRNA dynamics and cytokine profile in governing CD44v6/Nanog/PTEN axis in oral cancer: modulating the master regulators

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Tumor Biology

Abstract

Late diagnosis, low therapeutic response, and metastasis are accountable for poor 5-year survival rate of OSCC. These failures are attributed to the existence of “cancer stem cell (CSC)” subpopulation. Hence, it is necessary to identify and understand the mechanism of CSCs in tumor development, metastasis, and chemotherapeutic response. Propelling evidences suggest that microRNA (miRNA)-mediated regulation and cytokines of tumor microenvironment have the ability to modulate CSC signalling pathway; however, their exact mechanism needs to be elucidated. Thus, in this study, we characterized CSC markers and highlighted the miRNA dynamics and cytokine profile regulating these CSCs in a pathway-dependent manner. Our results demonstrated CD44+ subpopulation as tumor-initiating cells with self-renewal capability, tumorigenic growth potential and intrinsic chemoresistance. These tumors exhibited increased expression of CSC markers (CD44v3, CD44v6, Nanog, and Bmi1) and significantly reduced expression of PTEN and ATM in OSCC patients. Pathway analysis of these CSC markers demonstrated a prospective pathway regulated by miRNA and cytokine network. On analyzing these modulators, we observed decreased expression of miRNA542-3p, miRNA34a and miRNA9, and significant upregulation of miRNA21, thus forming an unexplored axis. Cytokine profiling revealed significantly increased levels of IL-6 and IL-8 compared to normals and demonstrated their strong association with CD44v6. Collectively, this study indicates that miR5423p and miR34a targets the CD44v6-Nanog-PTEN axis, thus playing a vital role in regulating the CSC properties. Furthermore, we speculate an impinging role of cytokines IL-6 and IL-8 in regulating this CSC-mediated pathway which can have prognostic and therapeutic implications.

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References

  1. Patel SS, Shah KA, Shah MJ, Kothari KC, Rawal RM. Cancer stem cells and stemness markers in oral squamous cell carcinomas. Asian Pac J Cancer Prev. 2014;15:8549–56.

    Article  PubMed  Google Scholar 

  2. McCullough MJ, Prasad G, Farah C. Oral mucosal malignancy and potentially malignant lesions: an update on the epidemiology, risk factors, diagnosis and management. Aust Dent J. 2010;55:61–5.

    Article  PubMed  Google Scholar 

  3. Mitra D, Malkoski SP, Wang XJ. Cancer stem cells in head and neck cancer. Cancers (Basel). 2011;3:415–27.

    Article  Google Scholar 

  4. Chikamatsu K, Ishii H, Takahashi G, Okamoto A, Moriyama M, Sakakura K, et al. Resistance to apoptosis-inducing stimuli in CD44+ head and neck squamous cell carcinoma cells. Head Neck. 2012;34:336–43.

    Article  PubMed  Google Scholar 

  5. Patel S, Shah K, Mirza S, Daga A, Rawal R. Epigenetic regulators governing cancer stem cells and epithelial-mesenchymal transition in oral squamous cell carcinoma. Curr Stem Cell Res Ther. 2015;10:140–52.

    Article  CAS  PubMed  Google Scholar 

  6. Prince ME, Sivanandan R, Kaczorowski A, Wolf GT, Kaplan MJ, Dalerba P, et al. Identification of a subpopulation of cells with cancer stem cell properties in head and neck squamous cell carcinoma. Proc Natl Acad Sci USA. 2007;104:973–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Bourguignon LY, Wong G, Earle C, Chen L. Hyaluronan-CD44v3 interaction with Oct4-Sox2-Nanog promotes miR-302 expression leading to self-renewal, clonal formation, and cisplatin resistance in cancer stem cells from head and neck squamous cell carcinoma. J Biol Chem. 2012;287:32800–24.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Paranjape AN, Balaji SA, Mandal T, Krushik EV, Nagaraj P, Mukherjee G, et al. Bmi1 regulates self-renewal and epithelial to mesenchymal transition in breast cancer cells through Nanog. BMC Cancer. 2014;14:785.

    Article  PubMed  PubMed Central  Google Scholar 

  9. Costea DE, Tsinkalovsky O, Vintermyr OK, Johannessen AC, Mackenzie IC. Cancer stem cells—new and potentially important targets for the therapy of oral squamous cell carcinoma. Oral Dis. 2006;12:443–54.

    Article  CAS  PubMed  Google Scholar 

  10. Clevers H. The cancer stem cell: premises, promises and challenges. Nat Med. 2011;17:313–9.

    Article  CAS  PubMed  Google Scholar 

  11. Shah A, Patel S, Pathak J, Swain N, Kumar S. The evolving concepts of cancer stem cells in head and neck squamous cell carcinoma. Scientific World Journal. 2014;2014:842491.

    PubMed  PubMed Central  Google Scholar 

  12. Braakhuis B, Leemans C, Brakenhoff RH. Expanding fields of genetically altered cells in head and neck squamous carcinogenesis. Semin Cancer Biol. 2005;15:113–20.

    Article  PubMed  Google Scholar 

  13. Golestaneh AF, Atashi A, Langroudi L, Shafiee A, Ghaemi N, Soleimani M. miRNAs expressed differently in cancer stem cells and cancer cells of human gastric cancer cell line MKN-45. Cell Biochem Funct. 2012;30:411–8.

    Article  CAS  PubMed  Google Scholar 

  14. Kashyap V, Rezende NC, Scotland KB, Shaffer SM, Persson JL, Gudas LJ, et al. Regulation of stem cell pluripotency and differentiation involves a mutual regulatory circuit of the NANOG, OCT4, and SOX2 pluripotency transcription factors with polycomb repressive complexes and stem cell microRNAs. Stem Cells Dev. 2009;18:1093–108.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Van der Horst G, Bos L, van der Pluijm G. Epithelial plasticity, cancer stem cells, and the tumor-supportive stroma in bladder carcinoma. Mol Cancer Res. 2012;10:995–1009.

    Article  PubMed  Google Scholar 

  16. Korkaya H, Liu S, Wicha MS. Breast cancer stem cells, cytokine networks, and the tumor microenvironment. J Clin Invest. 2011;121:3804–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Yang CH, Wang HL, Lin YS, Kumar KP, Lin HC, Chang CJ, et al. Identification of CD24 as a cancer stem cell marker in human nasopharyngeal carcinoma. PLoS One. 2014;9:e99412.

    Article  PubMed  PubMed Central  Google Scholar 

  18. Han J, Fujisawa T, Husain SR, Puri RK. Identification and characterization of cancer stem cells in human head and neck squamous cell carcinoma. BMC Cancer. 2014;14:173.

    Article  PubMed  PubMed Central  Google Scholar 

  19. Ghuwalewala S, Ghatak D, Das P, Dey S, Sarkar S, Alam N, et al. CD44(high)CD24(low) molecular signature determines the cancer stem cell and EMT phenotype in oral squamous cell carcinoma. Stem Cell Res. 2016;16:405–17.

    Article  CAS  PubMed  Google Scholar 

  20. Krishnamurthy S, Nör JE. Orosphere assay: a method for propagation of head and neck cancer stem cells. Head Neck. 2013;35:1015–21.

    Article  PubMed  Google Scholar 

  21. Nör C, Zhang Z, Warner KA, Bernardi L, Visioli F, Helman JI, et al. Cisplatin induces Bmi-1 and enhances the stem cell fraction in head and neck cancer. Neoplasia. 2014;16:137–46.

    Article  PubMed  PubMed Central  Google Scholar 

  22. Wang Y, Huang JW, Castella M, Huntsman DG, Taniguchi T. p53 is positively regulated by miR-542-3p. Cancer Res. 2014;74:3218–27.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Oneyama C, Morii E, Okuzaki D, Takahashi Y, Ikeda J, Wakabayashi N, et al. MicroRNA-mediated upregulation of integrin-linked kinase promotes Src-induced tumor progression. Oncogene. 2012;31:1623–35.

    Article  CAS  PubMed  Google Scholar 

  24. Misso G, Di Martino MT, De Rosa G, Farooqi AA, Lombardi A, Campani V, et al. Mir-34: a new weapon against cancer? Mol Ther Nucleic Acids. 2014;3:e194.

    Article  CAS  PubMed  Google Scholar 

  25. Yang Z, Fang S, Di Y, Ying W, Tan Y, Gu W. Modulation of NF-kB/miR-21/PTEN pathway sensitizes non-small cell lung cancer to cisplatin. PLoS One. 2015;10:e0121547.

    Article  PubMed  PubMed Central  Google Scholar 

  26. Zhang J, Jia J, Zhao L, Li X, Xie Q, Chen X, et al. Down-regulation of microRNA-9 leads to activation of IL-6/Jak/STAT3 pathway through directly targeting IL-6 in HeLa cell. Mol Carcinog. 2016;55:732–42.

    Article  PubMed  Google Scholar 

  27. St John MA, Li Y, Zhou X, Denny P, Ho CM, Montemagno C, et al. Interleukin 6 and interleukin 8 as potential biomarkers for oral cavity and oropharyngeal squamous cell carcinoma. Arch Otolaryngol Head Neck Surg. 2004;130:929–35.

    Article  PubMed  Google Scholar 

  28. Sansone P, Storci G, Tavolari S, Guarnieri T, Giovannini C, Taffurelli M, et al. IL-6 triggers malignant features in mammospheres from human ductal breast carcinoma and normal mammary gland. J Clin Invest. 2007;117:3988–4002.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. He Q, Liu Z, Zhao T, Zhao L, Zhou X, Wang A. Bmi1 drives stem-like properties and is associated with migration, invasion, and poor prognosis in tongue squamous cell carcinoma. Int J Biol Sci. 2015;11:1–10.

    Article  PubMed  PubMed Central  Google Scholar 

  30. Yu MA, Kiang A, Wang-Rodriguez J, Rahimy E, Haas M, Yu V, et al. Nicotine promotes acquisition of stem cell and epithelial-to-mesenchymal properties in head and neck squamous cell carcinoma. PLoS One. 2012;7:e51967.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgment

We thank Dr. Hemangini Vora (Head of Immunohistochemistry Division) for their excellent technical assistance for flow cytometry analysis. We are grateful to The Gujarat Cancer Society for their financial assistance.

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Authors and Affiliations

  1. Division of Medicinal Chemistry & Pharmacogenomics, Department of Cancer Biology, The Gujarat Cancer & Research Institute, Ahmedabad, Gujarat, 380016, India

    Shanaya Patel & Rakesh Rawal

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  1. Shanaya Patel
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  2. Rakesh Rawal
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Correspondence to Rakesh Rawal.

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This study was approved by the Institutional Review Board and Ethics Committee at The Gujarat Cancer Research Institute.

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Patel, S., Rawal, R. Role of miRNA dynamics and cytokine profile in governing CD44v6/Nanog/PTEN axis in oral cancer: modulating the master regulators. Tumor Biol. 37, 14565–14575 (2016). https://doi.org/10.1007/s13277-016-5289-2

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  • DOI: https://doi.org/10.1007/s13277-016-5289-2

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