Skip to main content

CD44-positive cancer stem cells from oral squamous cell carcinoma exhibit reduced proliferation and stemness gene expression upon adipogenic induction


We proposed to assess adipogenic differentiation and its effect on the proliferation and stemness markers in CD44 + OSCC CSCs. D44 + CSCs were sorted by magnetic sorting from the single-cell suspension of the OSCC tumor. Adipogenic differentiation was induced by an adipogenic induction medium. Lipid droplet formation was confirmed by oil red O staining. The expression of the cell surface marker was analyzed by flow cytometry. Real-time qPCR was performed to examine the gene expression activity. CD44 + OSCC CSCs can differentiate into adipocytes and adipogenesis in these cells decrease their proliferation and stemness gene expression. Adipogenic induction can make the cancer stem cells from OSCC tumors lose their stemness potential. Oral cancer, especially OSCC, is a huge burden worldwide. Similar to other stem cells, cancer stem cells can differentiate into other lineage cells. Our study shows that the proliferation and stemness gene expression in the CSCs from OSCC tumors can be thwarted by inducing them to differentiate into adipocytes, which could be advantageous to find out new clinical approaches in the treatment of cancers, like OSCC.

Graphical abstract

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4


  1. Montero PH, Patel SG. Cancer of the oral cavity. Surg Oncol Clin N Am. 2015;24:491–508.

    Article  Google Scholar 

  2. Bhutia SK, Naik PP, Praharaj PP, Panigrahi DP, Bhol CS, Mahapatra KK, et al. Identification and characterization of stem cells in oral cancer. Methods Mol Biol. 2019;2002:129–39.

    Article  CAS  Google Scholar 

  3. Naik PP, Das DN, Panda PK, Mukhopadhyay S, Sinha N, Praharaj PP, et al. Implications of cancer stem cells in developing therapeutic resistance in oral cancer. Oral Oncol. 2016;62:122–35.

    Article  CAS  Google Scholar 

  4. Phi LTH, Sari IN, Yang Y-G, Lee S-H, Jun N, Kim KS, et al. Cancer stem cells (CSCs) in drug resistance and their therapeutic implications in cancer treatment. Stem Cells Int. 2018;2018:5416923.

    Article  Google Scholar 

  5. Silva Galbiatti-Dias AL, Fernandes GMM, Castanhole-Nunes MMU, Hidalgo LF, Nascimento Filho CHV, Kawasaki-Oyama RS, et al. Relationship between CD44high/CD133high/CD117high cancer stem cells phenotype and Cetuximab and Paclitaxel treatment response in head and neck cancer cell lines. Am J Cancer Res. 2018;8:1633–41.

    PubMed  PubMed Central  Google Scholar 

  6. 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  Google Scholar 

  7. Mohanta S, Sekhar Khora S, Suresh A. Cancer stem cell based molecular predictors of tumor recurrence in oral squamous cell carcinoma. Arch Oral Biol. 2019;99:92–106.

    Article  CAS  Google Scholar 

  8. Noto Z, Yoshida T, Okabe M, Koike C, Fathy M, Tsuno H, et al. CD44 and SSEA-4 positive cells in an oral cancer cell line HSC-4 possess cancer stem-like cell characteristics. Oral Oncol. 2013;49:787–95.

    Article  CAS  Google Scholar 

  9. Shang H-G, Yu H-L, Ma X-N, Xu X. Multidrug resistance and tumor-initiating capacity of oral cancer stem cells. J BUON. 2016;21:461–5.

    PubMed  Google Scholar 

  10. Visvader JE, Lindeman GJ. Cancer stem cells in solid tumours: accumulating evidence and unresolved questions. Nat Rev Cancer. 2008;8:755–68.

    Article  CAS  Google Scholar 

  11. Gong L, Yan Q, Zhang Y, Fang X, Liu B, Guan X. Cancer cell reprogramming: a promising therapy converting malignancy to benignity. Cancer Commun (London, England). 2019;39:48.

    Article  Google Scholar 

  12. Xu B, O’Donnell M, O’Donnell J, Yu J, Zhang Y, Sartor MA, et al. Adipogenic differentiation of thyroid cancer cells through the Pax8-PPARγ fusion protein is regulated by thyroid transcription factor 1 (TTF-1). J Biol Chem. 2016;291:19274–86.

    Article  CAS  Google Scholar 

  13. Nassar D, Blanpain C. Cancer stem cells: basic concepts and therapeutic implications. Annu Rev Pathol Mech Dis. 2016;11:47–76.

    Article  CAS  Google Scholar 

  14. Takahashi N, Nobusue H, Shimizu T, Sugihara E, Yamaguchi-Iwai S, Onishi N, et al. ROCK inhibition induces terminal adipocyte differentiation and suppresses tumorigenesis in chemoresistant osteosarcoma cells. Cancer Res. 2019;79:3088–99.

    Article  CAS  Google Scholar 

  15. Kim YJ, Yu DB, Kim M, Choi Y-L. Adipogenesis induces growth inhibition of dedifferentiated liposarcoma. Cancer Sci. 2019;110:2676–83.

    Article  CAS  Google Scholar 

  16. Ma X, Zhao T, Ouyang T, Xin S, Ma Y, Chang M. Propranolol enhanced adipogenesis instead of induction of apoptosis of hemangiomas stem cells. Int J Clin Exp Pathol. 2014;7:3809–17.

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Choi S, Myers JN. Molecular pathogenesis of oral squamous cell carcinoma: implications for therapy. J Dent Res. 2008;87:14–32.

    Article  CAS  Google Scholar 

  18. Markopoulos AK. Current Aspects on Oral Squamous Cell Carcinoma. Open Dent J [Internet]. 2012;6:126–30. Available from:

  19. Jadhav KB, Gupta N. Clinicopathological prognostic implicators of oral squamous cell carcinoma: need to understand and revise. N Am J Med Sci. 2013;5:671–9.

    Article  Google Scholar 

  20. Liu S-A, Wang C-C, Jiang R-S, Lee F-Y, Lin W-J, Lin J-C. Pathological features and their prognostic impacts on oral cavity cancer patients among different subsites – A singe institute’s experience in Taiwan. Sci Rep. 2017;7:7451.

    Article  Google Scholar 

  21. Almangush A, Bello IO, Coletta RD, Mäkitie AA, Mäkinen LK, Kauppila JH, et al. For early-stage oral tongue cancer, depth of invasion and worst pattern of invasion are the strongest pathological predictors for locoregional recurrence and mortality. Virchows Arch. 2015;467:39–46.

    Article  Google Scholar 

  22. Zhang C-L, Huang T, Wu B-L, He W-X, Liu D. Stem cells in cancer therapy: opportunities and challenges. Oncotarget. 2017;8:75756–66.

    Article  Google Scholar 

  23. Bajaj J, Diaz E, Reya T. Stem cells in cancer initiation and progression. J Cell Biol. 2020.

    Article  PubMed  Google Scholar 

  24. Mackenzie IC. Growth of malignant oral epithelial stem cells after seeding into organotypical cultures of normal mucosa. J Oral Pathol Med. 2004;33:71–8.

    Article  Google Scholar 

  25. Okamoto A, Chikamatsu K, Sakakura K, Hatsushika K, Takahashi G, Masuyama K. Expansion and characterization of cancer stem-like cells in squamous cell carcinoma of the head and neck. Oral Oncol. 2009;45:633–9.

    Article  CAS  Google Scholar 

  26. Chiou S-H, Yu C-C, Huang C-Y, Lin S-C, Liu C-J, Tsai T-H, et al. Positive correlations of Oct-4 and Nanog in oral cancer stem-like cells and high-grade oral squamous cell carcinoma. Clin Cancer Res. 2008;14:4085–95.

    Article  CAS  Google Scholar 

  27. Thavarool SB, Muttath G, Nayanar S, Duraisamy K, Bhat P, Shringarpure K, et al. Improved survival among oral cancer patients: findings from a retrospective study at a tertiary care cancer centre in rural Kerala. India World J Surg Oncol. 2019;17:15.

    Article  Google Scholar 

  28. Shekhani MT, Jayanthy A-S, Maddodi N, Setaluri V. Cancer stem cells and tumor transdifferentiation: implications for novel therapeutic strategies. Am J Stem Cells. 2013;2:52–61.

    CAS  PubMed  PubMed Central  Google Scholar 

  29. Castaigne S, Chomienne C, Daniel MT, Ballerini P, Berger R, Fenaux P, et al. All-trans retinoic acid as a differentiation therapy for acute promyelocytic leukemia. I Clinical results Blood. 1990;76:1704–9.

    CAS  PubMed  Google Scholar 

  30. Yan Y, Li Z, Xu X, Chen C, Wei W, Fan M, et al. All-trans retinoic acids induce differentiation and sensitize a radioresistant breast cancer cells to chemotherapy. BMC Complement Altern Med. 2016;16:113.

    Article  Google Scholar 

  31. Baillie R, Tan ST, Itinteang T. Cancer Stem Cells in Oral Cavity Squamous Cell Carcinoma: A Review. Front Oncol. 2017;7:112.

    Article  Google Scholar 

  32. Oliveira LR, Castilho-Fernandes A, Oliveira-Costa JP, Soares FA, Zucoloto S, Ribeiro-Silva A. CD44+/CD133+ immunophenotype and matrix metalloproteinase-9: influence on prognosis in early-stage oral squamous cell carcinoma. Head Neck. 2014;36:1718–26.

    Article  Google Scholar 

  33. Dobson ME, Diallo-Krou E, Grachtchouk V, Yu J, Colby LA, Wilkinson JE, et al. Pioglitazone induces a proadipogenic antitumor response in mice with PAX8-PPARgamma fusion protein thyroid carcinoma. Endocrinology. 2011;152:4455–65.

    Article  CAS  Google Scholar 

  34. Basu-Roy U, Han E, Rattanakorn K, Gadi A, Verma N, Maurizi G, et al. PPARγ agonists promote differentiation of cancer stem cells by restraining YAP transcriptional activity. Oncotarget. 2016;7:60954–70.

    Article  Google Scholar 

  35. Hallenborg P, Feddersen S, Madsen L, Kristiansen K. The tumor suppressors pRB and p53 as regulators of adipocyte differentiation and function. Expert Opin Ther Targets. 2009;13:235–46.

    Article  CAS  Google Scholar 

  36. Rosen ED, Hsu C-H, Wang X, Sakai S, Freeman MW, Gonzalez FJ, et al. C/EBPalpha induces adipogenesis through PPARgamma: a unified pathway. Genes Dev. 2002;16:22–6.

    Article  CAS  Google Scholar 

  37. Avilion AA. Multipotent cell lineages in early mouse development depend on SOX2 function. Genes Dev. 2003;17:126–40.

    Article  CAS  Google Scholar 

  38. Wefers C, Schreibelt G, Massuger LFAG, de Vries IJM, Torensma R. Immune curbing of cancer stem cells by CTLs directed to NANOG. Front Immunol. 2018.

    Article  PubMed  PubMed Central  Google Scholar 

  39. Takahashi K, Yamanaka S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell. 2006;126:663–76.

    Article  CAS  Google Scholar 

Download references


The present work was funded by the Taif University Researchers Supporting Program (Project number: TURSP-2020/151), Taif University, Saudi Arabia.

Author information

Authors and Affiliations


Corresponding author

Correspondence to Shankargouda Patil.

Ethics declarations

Conflict of interest

The authors declare no conflict of interest.

Ethical approval

The study was conducted according to the guidelines of the Declaration of Helsinki. The research protocol was approved by Scientific Research, College of Dentistry, Jazan University. Reference Number: CODJU-19212.

Informed consent

Not Applicable.

Additional information

Publisher's Note

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

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Patil, S., Al-Brakati, A., Abidi, N.H. et al. CD44-positive cancer stem cells from oral squamous cell carcinoma exhibit reduced proliferation and stemness gene expression upon adipogenic induction. Med Oncol 39, 23 (2022).

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI:


  • Adipogenic differentiation
  • Cancer stem cells
  • CD44
  • OSCC
  • Stemness