Clinical Oral Investigations

, Volume 20, Issue 1, pp 43–56

Prognostic role of Oct4, CD44 and c-Myc in radio–chemo-resistant oral cancer patients and their tumourigenic potential in immunodeficient mice

  • Sharada Sawant
  • Ravindran Gokulan
  • Harsh Dongre
  • Milind Vaidya
  • Devendra Chaukar
  • Kumar Prabhash
  • Arvind Ingle
  • Shriya Joshi
  • Prerana Dange
  • Shreyas Joshi
  • Archana Kumari Singh
  • Vidhi Makani
  • Shilpi Sharma
  • Ashok Jeyaram
  • Shubhada Kane
  • Anil D’Cruz
Original Article



In the present study, we have investigated the prognostic value of known stem cell-associated molecules such as Oct4, CD44 and c-Myc in patients with oral SCC who had received post-surgery radio- and/or chemotherapy.

Materials and methods

Immunohistochemistry was performed to analyse the expression of Oct4, CD44 and c-Myc in 87 tumour tissues, and the expression profile obtained was correlated with clinicopathological parameters of the patients with oral cancer. Tumourigenic potential of these molecules was also evaluated by in vivo studies.


Our results showed significant correlation of Oct4 (OS, p = 0.003; DFS, p = 0.001) and c-Myc (OS, p = 0.01; DFS, p = 0.03) with overall survival and disease-free survival independently. Furthermore, all the three markers in combinations of two markers each, i.e. Oct4 + CD44 (OS, p = 0.003; DFS, p = 0.001), Oct4 + c-Myc (OS, p = 0.0001; DFS, p = 0.0001), CD44 + c-Myc (OS, p = 0.008; DFS, p = 0.02) and in combinations of three markers each, i.e. Oct4 + CD44 + c-Myc (OS, p = 0.0001; DFS, p = 0.0001) also significantly correlated with overall survival and disease-free survival. Univariate and multivariate analyses further established the independent prognostic value of Oct4. Oct4-, CD44- and c-Myc-enriched populations independently induced sarcomatoid carcinomas whereas primary keratinocytes developed poorly differentiated carcinomas in immunodeficient mice.


Oct4 and c-Myc independently as well as in combination with CD44 might be useful for the prediction of local recurrence and poor survival of patients with oral cancer which is the novel finding of this study.

Clinical relevance

Oct4, c-Myc and CD44 can be used to predict local recurrence and the outcome of treatment in oral cancer patients. In addition, these molecules may find use as molecular targets for effective therapy.


Oral cancer Recurrence Cancer stem cells Oct4 CD44 Prognosis 


  1. 1.
    Warnakulasuriya S (2009) Global epidemiology of oral and oropharyngeal cancer. Oral Oncol 45(4–5):309–316. doi:10.1016/j.oraloncology.2008.06.002 PubMedCrossRefGoogle Scholar
  2. 2.
    Jemal A, Siegel R, Ward E, Hao Y, Xu J, Murray T, Thun MJ (2008) Cancer statistics, 2008. CA Cancer J Clin 58(2):71–96. doi:10.3322/ca.2007.0010 PubMedCrossRefGoogle Scholar
  3. 3.
    da Silva SD, Hier M, Mlynarek A, Kowalski LP, Alaoui-Jamali MA (2012) Recurrent oral cancer: current and emerging therapeutic approaches. Front Pharmacol 3:149. doi:10.3389/fphar.2012.00149 PubMedPubMedCentralCrossRefGoogle Scholar
  4. 4.
    Prince ME, Sivanandan R, Kaczorowski A, Wolf GT, Kaplan MJ, Dalerba P, Weissman IL, Clarke MF, Ailles LE (2007) Identification of a subpopulation of cells with cancer stem cell properties in head and neck squamous cell carcinoma. Proc Natl Acad Sci U S A 104(3):973–978. doi:10.1073/pnas.0610117104 PubMedPubMedCentralCrossRefGoogle Scholar
  5. 5.
    Pries R, Witrkopf N, Trenkle T, Nitsch SM, Wollenberg B (2008) Potential stem cell marker CD44 is constitutively expressed in permanent cell lines of head and neck cancer. In Vivo (Athens, Greece) 22(1):89–92Google Scholar
  6. 6.
    Huang CF, Xu XR, Wu TF, Sun ZJ, Zhang WF (2014) Correlation of ALDH1, CD44, OCT4 and SOX2 in tongue squamous cell carcinoma and their association with disease progression and prognosis. J Oral Pathol Med Off Publ Int Ass Oral Pathol Am Acad Oral Pathol 43(7):492–498. doi:10.1111/jop.12159 Google Scholar
  7. 7.
    Lee J, Kim HK, Rho JY, Han YM, Kim J (2006) The human OCT-4 isoforms differ in their ability to confer self-renewal. J Biol Chem 281(44):33554–33565. doi:10.1074/jbc.M603937200 PubMedCrossRefGoogle Scholar
  8. 8.
    Pesce M, Scholer HR (2001) Oct-4: gatekeeper in the beginnings of mammalian development. Stem Cells (Dayton, Ohio) 19(4):271–278. doi:10.1634/stemcells.19-4-271 CrossRefGoogle Scholar
  9. 9.
    Tai MH, Chang CC, Kiupel M, Webster JD, Olson LK, Trosko JE (2005) Oct4 expression in adult human stem cells: evidence in support of the stem cell theory of carcinogenesis. Carcinogenesis 26(2):495–502. doi:10.1093/carcin/bgh321 PubMedCrossRefGoogle Scholar
  10. 10.
    Chiou SH, Yu CC, Huang CY, Lin SC, Liu CJ, Tsai TH, Chou SH, Chien CS, Ku HH, Lo JF (2008) Positive correlations of Oct-4 and Nanog in oral cancer stem-like cells and high-grade oral squamous cell carcinoma. Clin Cancer Res Off J Am Assoc Cancer Res 14(13):4085–4095. doi:10.1158/1078-0432.ccr-07-4404 CrossRefGoogle Scholar
  11. 11.
    Ravindran G, Sawant SS, Hague A, Kingsley K, Devaraj H (2014) Association of differential beta-catenin expression with Oct-4 and Nanog in oral squamous cell carcinoma and their correlation with clinicopathological factors and prognosis. Head Neck. doi:10.1002/hed.23699 PubMedGoogle Scholar
  12. 12.
    Tsai LL, Yu CC, Chang YC, Yu CH, Chou MY (2011) Markedly increased Oct4 and Nanog expression correlates with cisplatin resistance in oral squamous cell carcinoma. J Oral Pathol Med Off Publ Int Ass Oral Pathol Am Acad Oral Pathol 40(8):621–628. doi:10.1111/j.1600-0714.2011.01015.x Google Scholar
  13. 13.
    Beltran AS, Rivenbark AG, Richardson BT, Yuan X, Quian H, Hunt JP, Zimmerman E, Graves LM, Blancafort P (2011) Generation of tumor-initiating cells by exogenous delivery of OCT4 transcription factor. Breast Cancer Res BCR 13(5):R94. doi:10.1186/bcr3019 PubMedCrossRefGoogle Scholar
  14. 14.
    Soukka T, Salmi M, Joensuu H, Hakkinen L, Sointu P, Koulu L, Kalimo K, Klemi P, Grenman R, Jalkanen S (1997) Regulation of CD44v6-containing isoforms during proliferation of normal and malignant epithelial cells. Cancer Res 57(11):2281–2289PubMedGoogle Scholar
  15. 15.
    Zeilstra J, Joosten SP, Dokter M, Verwiel E, Spaargaren M, Pals ST (2008) Deletion of the WNT target and cancer stem cell marker CD44 in Apc(Min/+) mice attenuates intestinal tumorigenesis. Cancer Res 68(10):3655–3661. doi:10.1158/0008-5472.can-07-2940 PubMedCrossRefGoogle Scholar
  16. 16.
    Takaishi S, Okumura T, Tu S, Wang SS, Shibata W, Vigneshwaran R, Gordon SA, Shimada Y, Wang TC (2009) Identification of gastric cancer stem cells using the cell surface marker CD44. Stem Cells (Dayton, Ohio) 27(5):1006–1020. doi:10.1002/stem.30 CrossRefGoogle Scholar
  17. 17.
    Ue T, Yokozaki H, Kagai K, Higashikawa K, Yasui W, Sugiyama M, Tahara E, Ishikawa T (1998) Reduced expression of the CD44 variant exons in oral squamous cell carcinoma and its relationship to metastasis. J Oral Pathol Med Off Publ Int Ass Oral Pathol Am Acad Oral Pathol 27(5):197–201Google Scholar
  18. 18.
    Monroe MM, Anderson EC, Clayburgh DR, Wong MH (2011) Cancer stem cells in head and neck squamous cell carcinoma. J Oncol 2011:762780. doi:10.1155/2011/762780 PubMedPubMedCentralCrossRefGoogle Scholar
  19. 19.
    Masuda M, Kuratomi Y, Shiratsuchi H, Nakashima T, Naonobu K, Komiyama S (2000) Decreased CD44H expression in early-stage tongue carcinoma associates with late nodal metastases following interstitial brachytherapy. Head Neck 22(7):662–665PubMedCrossRefGoogle Scholar
  20. 20.
    Oliveira DT, Sherriff M, Odell EW (1998) Expression of CD44 variant exons by primary and metastatic oral squamous carcinomas. J Oral Pathol Med Off Publ Int Ass Oral Pathol Am Acad Oral Pathol 27(7):303–307Google Scholar
  21. 21.
    Gonzalez-Moles MA, Bravo M, Ruiz-Avila I, Esteban F, Bascones-Martinez A, Gonzalez-Moles S (2004) Adhesion molecule CD44 expression in non-tumour epithelium adjacent to tongue cancer. Oral Oncol 40(3):281–286PubMedCrossRefGoogle Scholar
  22. 22.
    Kunishi M, Kayada Y, Yoshiga K (1997) Down-regulated expression of CD44 variant 6 in oral squamous cell carcinomas and its relationship to regional lymph node metastasis. Int J Oral Maxillofac Surg 26(4):280–283PubMedCrossRefGoogle Scholar
  23. 23.
    Lindquist D, Ahrlund-Richter A, Tarjan M, Tot T, Dalianis T (2012) Intense CD44 expression is a negative prognostic factor in tonsillar and base of tongue cancer. Anticancer Res 32(1):153–161PubMedGoogle Scholar
  24. 24.
    Abdulmajeed AA, Dalley AJ, Farah CS (2013) Putative cancer stem cell marker expression in oral epithelial dysplasia and squamous cell carcinoma. J Oral Pathol Med Off Publ Int Ass Oral Pathol Am Acad Oral Pathol 42(10):755–760. doi:10.1111/jop.12073 Google Scholar
  25. 25.
    Bankfalvi A, Krassort M, Buchwalow IB, Vegh A, Felszeghy E, Piffko J (2002) Gains and losses of adhesion molecules (CD44, E-cadherin, and beta-catenin) during oral carcinogenesis and tumour progression. J Pathol 198(3):343–351. doi:10.1002/path.1204 PubMedCrossRefGoogle Scholar
  26. 26.
    Cichy J, Pure E (2003) The liberation of CD44. J Cell Biol 161(5):839–843. doi:10.1083/jcb.200302098 PubMedPubMedCentralCrossRefGoogle Scholar
  27. 27.
    Rautava J, Soukka T, Inki P, Leimola-Virtanen R, Saloniemi I, Happonen RP, Heikinheimo K (2003) CD44v6 in developing, dysplastic and malignant oral epithelia. Oral Oncol 39(4):373–379PubMedCrossRefGoogle Scholar
  28. 28.
    Du L, Wang H, He L, Zhang J, Ni B, Wang X, Jin H, Cahuzac N, Mehrpour M, Lu Y, Chen Q (2008) CD44 is of functional importance for colorectal cancer stem cells. Clin Cancer Res Off J Am Assoc Cancer Res 14(21):6751–6760. doi:10.1158/1078-0432.ccr-08-1034 CrossRefGoogle Scholar
  29. 29.
    Dang CV (2012) MYC on the path to cancer. Cell 149(1):22–35. doi:10.1016/j.cell.2012.03.003 PubMedPubMedCentralCrossRefGoogle Scholar
  30. 30.
    Klinakis A, Szabolcs M, Politi K, Kiaris H, Artavanis-Tsakonas S, Efstratiadis A (2006) Myc is a Notch1 transcriptional target and a requisite for Notch1-induced mammary tumorigenesis in mice. Proc Natl Acad Sci U S A 103(24):9262–9267. doi:10.1073/pnas.0603371103 PubMedPubMedCentralCrossRefGoogle Scholar
  31. 31.
    Shao Y, Qu Y, Dang S, Yao B, Ji M (2013) MiR-145 inhibits oral squamous cell carcinoma (OSCC) cell growth by targeting c-Myc and Cdk6. Cancer Cell Int 13(1):51. doi:10.1186/1475-2867-13-51 PubMedPubMedCentralCrossRefGoogle Scholar
  32. 32.
    Gandarillas A, Watt FM (1997) c-Myc promotes differentiation of human epidermal stem cells. Genes Dev 11(21):2869–2882PubMedPubMedCentralCrossRefGoogle Scholar
  33. 33.
    Srinivasan M, Jewell SD (2001) Quantitative estimation of PCNA, c-myc, EGFR and TGF-alpha in oral submucous fibrosis—an immunohistochemical study. Oral Oncol 37(5):461–467PubMedCrossRefGoogle Scholar
  34. 34.
    Segura S, Rozas-Munoz E, Toll A, Martin-Ezquerra G, Masferrer E, Espinet B, Rodriguez M, Baro T, Barranco C, Pujol RM (2013) Evaluation of MYC status in oral lichen planus in patients with progression to oral squamous cell carcinoma. Br J Dermatol 169(1):106–114. doi:10.1111/bjd.12303 PubMedCrossRefGoogle Scholar
  35. 35.
    Shah NG, Trivedi TI, Tankshali RA, Goswami JV, Jetly DH, Shukla SN, Shah PM, Verma RJ (2009) Prognostic significance of molecular markers in oral squamous cell carcinoma: a multivariate analysis. Head Neck 31(12):1544–1556. doi:10.1002/hed.21126 PubMedCrossRefGoogle Scholar
  36. 36.
    Pindborg JJ (1997) Histological typing of cancer and precancer of the oral mucosa. Springer, BerlinCrossRefGoogle Scholar
  37. 37.
    Sawant SS, Chaukar DA, Joshi SS, Dange PP, Kannan S, Kane S, D’Cruz AK, Vaidya MM (2011) Prognostic value of tissue polypeptide antigen in oral squamous cell carcinoma. Oral Oncol 47(2):114–120. doi:10.1016/j.oraloncology.2010.11.005 PubMedCrossRefGoogle Scholar
  38. 38.
    Varghese F, Bukhari AB, Malhotra R, De A (2014) IHC Profiler: an open source plugin for the quantitative evaluation and automated scoring of immunohistochemistry images of human tissue samples. PLoS One 9(5), e96801. doi:10.1371/journal.pone.0096801 PubMedPubMedCentralCrossRefGoogle Scholar
  39. 39.
    Sawant SS, Vaidya M, Chaukar DA, Alam H, Dmello C, Gangadaran P, Kannan S, Kane S, Dange PP, Dey N, Ranganathan K, D’Cruz AK (2014) Clinical significance of aberrant vimentin expression in oral premalignant lesions and carcinomas. Oral Dis 20(5):453–465. doi:10.1111/odi.12151 PubMedCrossRefGoogle Scholar
  40. 40.
    Chinn SB, Darr OA, Peters RD, Prince ME (2012) The role of head and neck squamous cell carcinoma cancer stem cells in tumorigenesis, metastasis, and treatment failure. Front Endocrinol 3. doi:10.3389/fendo.2012.00090
  41. 41.
    Kosunen A, Pirinen R, Ropponen K, Pukkila M, Kellokoski J, Virtaniemi J, Sironen R, Juhola M, Kumpulainen E, Johansson R, Nuutinen J, Kosma VM (2007) CD44 expression and its relationship with MMP-9, clinicopathological factors and survival in oral squamous cell carcinoma. Oral Oncol 43(1):51–59. doi:10.1016/j.oraloncology.2006.01.003 PubMedCrossRefGoogle Scholar
  42. 42.
    Fonseca I, Pereira T, Rosa-Santos J, Soares J (2001) Expression of CD44 isoforms in squamous cell carcinoma of the border of the tongue: a correlation with histological grade, pattern of stromal invasion, and cell differentiation. J Surg Oncol 76(2):115–120PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Sharada Sawant
    • 1
  • Ravindran Gokulan
    • 2
  • Harsh Dongre
    • 1
  • Milind Vaidya
    • 1
  • Devendra Chaukar
    • 3
  • Kumar Prabhash
    • 4
  • Arvind Ingle
    • 5
  • Shriya Joshi
    • 1
  • Prerana Dange
    • 6
  • Shreyas Joshi
    • 7
  • Archana Kumari Singh
    • 1
  • Vidhi Makani
    • 8
  • Shilpi Sharma
    • 9
  • Ashok Jeyaram
    • 10
  • Shubhada Kane
    • 11
  • Anil D’Cruz
    • 9
  1. 1.Vaidya Laboratory, Advanced Centre for Treatment, Research and Education in Cancer (ACTREC)Tata Memorial CentreNavi MumbaiIndia
  2. 2.Department of Oral Biology and Biomedical Sciences, Faculty of DentistryUniversity of MalayaKuala LumpurMalaysia
  3. 3.Head and Neck UnitTata Memorial Hospital (TMH)MumbaiIndia
  4. 4.Medical Oncology UnitTata Memorial Hospital (TMH)MumbaiIndia
  5. 5.Laboratory Animal Facility, Advanced Centre for Treatment, Research and Education in Cancer (ACTREC)Tata Memorial CentreNavi MumbaiIndia
  6. 6.Sarin Laboratory, Advanced Centre for Treatment, Research and Education in Cancer (ACTREC)Tata Memorial CentreNavi MumbaiIndia
  7. 7.Department of BiologyUniversity of KentuckyLexingtonUSA
  8. 8.Laboratory of Chromatin Biology, Department of Biological SciencesIndian Institute of Science Education and ResearchBhopalIndia
  9. 9.Head and Neck UnitTata Memorial Hospital (TMH)MumbaiIndia
  10. 10.Microbiology LaboratoryCoimbatoreIndia
  11. 11.Pathology DepartmentTata Memorial Hospital (TMH)MumbaiIndia

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