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The stem cell markers expression CD44v6 and podoplanin in lip cancer: clinical significance

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Abstract

This study aimed to analyze the immunoexpression of cancer stem cell markers, CD44v6, and podoplanin in 91 patients with lip squamous cell carcinomas (LSCC). The immunostaining of podoplanin and CD44v6 was evaluated in ten high-power fields (× 400 magnification) at the invasive front of LSCC, using a semi-quantitative score method. Chi-square test or Fisher’s exact test was used to verify the association of podoplanin and CD44v6 expressions with clinicopathologic variables. Spearman’s correlation test was used to analyze the correlation between the two antibodies in lip cancer. Disease-free survival probabilities in 5 and 10 years were estimated according to the Kaplan-Meier method and compared using the log-rank test. The independent effects of the significant variables were analyzed by Cox proportional hazards regression model. A strong podoplanin expression was observed in the membrane and cytoplasm of most lip tumor cells, and this was inversely associated with locoregional recurrence (p = 0.028) and with histopathological grade of malignancy (p = 0.026). Additionally, CD44v6 immunostaining was strongly expressed in the membrane of tumor cells in 95.4% of the LSCC. Patients with strong membranous (p = 0.016) or strong cytoplasmic (p = 0.030) podoplanin-positive tumors resulted in significantly better disease-free survival than those who had podoplanin weak/negative tumors, confirming podoplanin expression as a favorable independent prognostic factor. Podoplanin and CD44v6 were strongly expressed by tumor cells and podoplanin immunoexpression can help to determine lip cancer patients with lower risk for disease recurrence.

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References

  1. Yanamoto S, Yamada S, Takahashi H, Naruse T, Matsushita Y, Ikeda H, Shiraishi T, Seki S, Fujita S, Ikeda T, Asahina I, Umeda M (2014) Expression of the cancer stem cell markers CD44v6 and ABCG2 in tongue cancer: effect of neoadjuvant chemotherapy on local recurrence. Int J Oncol 44(4):1153–1162. https://doi.org/10.3892/ijo.2014.2289

    Article  CAS  PubMed  Google Scholar 

  2. Athanassiou-Papaefthymiou M, Shkeir O, Kim D, Divi V, Matossian M, Owen JH, Czerwinski MJ, Papagerakis P, McHugh J, Bradford CR, Carey TE, Wolf GT, Prince ME, Papagerakis S (2014) Evaluation of CD44 variant expression in oral, head and neck squamous cell carcinomas using a triple approach and its clinical significance. Int J Immunopathol Pharmacol 27(3):337–349

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Sawant S, Gokulan R, Dongre H, Vaidya M, Chaukar D, Prabhash K, Ingle A, Joshi S, Dange P, Singh AK, Makani V, Sharma S, Jeyaram A, Kane S, D’Cruz A (2016) Prognostic role of Oct4, CD44 and c-Myc in radio-chemo-resistant oral cancer patients and their tumourigenic potential in immunodeficient mice. Clin Oral Investig 20(1):43–56. https://doi.org/10.1007/s00784-015-1476-610.1007/s00784-015-1476-6

    Article  PubMed  Google Scholar 

  4. Patel S, Rawal R (2016) Role of miRNA dynamics and cytokine profile in governing CD44v6/Nanog/PTEN axis in oral cancer: modulating the master regulators. Tumour Biol 37(11):14565–14575. https://doi.org/10.1007/s13277-016-5289-210.1007/s13277-016-5289-2

    Article  CAS  PubMed  Google Scholar 

  5. Baillie R, Tan ST, Itinteang T (2017) Cancer stem cells in oral cavity squamous cell carcinoma: a review. Front Oncol 7:112. https://doi.org/10.3389/fonc.2017.00112

    Article  PubMed  PubMed Central  Google Scholar 

  6. Mohanta S, Siddappa G, Valiyaveedan SG, Dodda Thimmasandra Ramanjanappa R, Das D, Pandian R, Khora SS, Kuriakose MA, Suresh A (2017) Cancer stem cell markers in patterning differentiation and in prognosis of oral squamous cell carcinoma. Tumour Biol 39(6):1010428317703656. https://doi.org/10.1177/1010428317703656

    Article  PubMed  Google Scholar 

  7. Boxberg M, Gotz C, Haidari S, Dorfner C, Jesinghaus M, Drecoll E, Boskov M, Wolff KD, Weichert W, Haller B, Kolk A (2018) Immunohistochemical expression of CD44 in oral squamous cell carcinoma in relation to histomorphologic parameters and clinicopathological factors. Histopathology. https://doi.org/10.1111/his.13496

  8. Vormittag L, Thurnher D, Geleff S, Pammer J, Heiduschka G, Brunner M, Grasl M, Erovic BM (2009) Co-expression of Bmi-1 and podoplanin predicts overall survival in patients with squamous cell carcinoma of the head and neck treated with radio(chemo)therapy. Int J Radiat Oncol Biol Phys 73(3):913–918. https://doi.org/10.1016/j.ijrobp.2008.10.040S0360-3016(08)03681-X

    Article  CAS  PubMed  Google Scholar 

  9. Habiba U, Hida K, Kitamura T, Matsuda AY, Higashino F, Ito YM, Ohiro Y, Totsuka Y, Shindoh M (2017) ALDH1 and podoplanin expression patterns predict the risk of malignant transformation in oral leukoplakia. Oncol Lett 13(1):321–328. https://doi.org/10.3892/ol.2016.5379OL-0-0-5379

    Article  CAS  PubMed  Google Scholar 

  10. Siar CH, Ishak I, Ng KH (2015) Podoplanin, E-cadherin, beta-catenin, and CD44v6 in recurrent ameloblastoma: their distribution patterns and relevance. J Oral Pathol Med 44(1):51–58. https://doi.org/10.1111/jop.12203

    Article  CAS  PubMed  Google Scholar 

  11. Dos Santos AA, Oliveira DT, Pereira MC, Faustino SE, Nonogaki S, Carvalho AL, Kowalski LP (2013) Podoplanin and VEGF-C immunoexpression in oral squamous cell carcinomas: prognostic significance. Anticancer Res 33(9):3969–3976

    Google Scholar 

  12. Caetano Ados S, Tjioe KC, Faustino SE, Hanemann JA, Belone Ade F, Soares CT, Oliveira DT (2013) Immunolocalization of podoplanin in benign odontogenic tumours with and without ectomesenchyme. Arch Oral Biol 58(4):408–415. https://doi.org/10.1016/j.archoralbio.2012.06.002S0003-9969(12)00186-0

    Article  PubMed  Google Scholar 

  13. Garcia AS, Tjioe KC, Nonogaki S, Soares FA, Soares CT, Lauris JRP, Oliveira DT (2014) Ezrin and podoplanin expressions are associated in invasion front of lip cancer. Head Neck Oncololy 3(1):6

    Google Scholar 

  14. Oliveira DT, de Santis LP, Assao A, Tjioe KC, Nonogaki S, Lauris JR, Soares FA (2014) The relationship between ezrin and podoplanin expressions in keratocystic odontogenic tumors. BMC Oral Health 14:150. https://doi.org/10.1186/1472-6831-14-1501472-6831-14-150

    Article  PubMed  PubMed Central  Google Scholar 

  15. Assao A, Nonogaki S, Lauris JR, Carvalho AL, Pinto CA, Soares FA, Kowalski LP, Oliveira DT (2016) Podoplanin, ezrin, and Rho-A proteins may have joint participation in tumor invasion of lip cancer. Clin Oral Investig. https://doi.org/10.1007/s00784-016-1956-310.1007/s00784-016-1956-3

  16. Barros FBA, Assao A, Garcia NG, Nonogaki S, Carvalho AL, Soares FA, Kowalski LP, Oliveira DT (2018) Moesin expression by tumor cells is an unfavorable prognostic biomarker for oral cancer. BMC Cancer 18(1):53. https://doi.org/10.1186/s12885-017-3914-0

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Costa YF, Tjioe KC, Nonogaki S, Soares FA, Lauris JR, Oliveira DT (2015) Are podoplanin and ezrin involved in the invasion process of the ameloblastomas? Eur J Histochem 59(1):2451. https://doi.org/10.4081/ejh.2015.2451

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Ozkul Y, Songu M, Imre A, Tunc E, Ozkul Z, Arslanoglu S, Pinar E, Onal K (2016) Early stage squamous cell carcinoma of the lower lip: predictive factors for recurrence. J Laryngol Otol 130(4):369–372. https://doi.org/10.1017/S0022215116000311S0022215116000311

    Article  CAS  PubMed  Google Scholar 

  19. Martin-Villar E, Scholl FG, Gamallo C, Yurrita MM, Munoz-Guerra M, Cruces J, Quintanilla M (2005) Characterization of human PA2.26 antigen (T1alpha-2, podoplanin), a small membrane mucin induced in oral squamous cell carcinomas. Int J Cancer 113(6):899–910. https://doi.org/10.1002/ijc.20656

    Article  CAS  PubMed  Google Scholar 

  20. Martin-Villar E, Megias D, Castel S, Yurrita MM, Vilaro S, Quintanilla M (2006) Podoplanin binds ERM proteins to activate RhoA and promote epithelial-mesenchymal transition. J Cell Sci 119(Pt 21):4541–4553. https://doi.org/10.1242/jcs.03218

    Article  CAS  PubMed  Google Scholar 

  21. Fernandez-Munoz B, Yurrita MM, Martin-Villar E, Carrasco-Ramirez P, Megias D, Renart J, Quintanilla M (2011) The transmembrane domain of podoplanin is required for its association with lipid rafts and the induction of epithelial-mesenchymal transition. Int J Biochem Cell Biol 43(6):886–896. https://doi.org/10.1016/j.biocel.2011.02.010S1357-2725(11)00066-5

    Article  CAS  PubMed  Google Scholar 

  22. Martin-Villar E, Fernandez-Munoz B, Parsons M, Yurrita MM, Megias D, Perez-Gomez E, Jones GE, Quintanilla M (2010) Podoplanin associates with CD44 to promote directional cell migration. Mol Biol Cell 21(24):4387–4399. https://doi.org/10.1091/mbc.E10-06-0489E10-06-0489

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Tsuneki M, Yamazaki M, Maruyama S, Cheng J, Saku T (2013) Podoplanin-mediated cell adhesion through extracellular matrix in oral squamous cell carcinoma. Lab Investig 93(8):921–932. https://doi.org/10.1038/labinvest.2013.86labinvest201386

    Article  CAS  Google Scholar 

  24. Yu YH, Morales J, Feng L, Lee JJ, El-Naggar AK, Vigneswaran N (2015) CD147 and Ki-67 overexpression confers poor prognosis in squamous cell carcinoma of oral tongue: a tissue microarray study. Oral Surg Oral Med Oral Pathol Oral Radiol 119(5):553–565. https://doi.org/10.1016/j.oooo.2014.12.022S2212-4403(14)01451-5

    Article  PubMed  PubMed Central  Google Scholar 

  25. Saghravanian N, Anvari K, Ghazi N, Memar B, Shahsavari M, Aghaee MA (2017) Expression of p63 and CD44 in oral squamous cell carcinoma and correlation with clinicopathological parameters. Arch Oral Biol 82:160–165. https://doi.org/10.1016/j.archoralbio.2017.06.011

    Article  CAS  PubMed  Google Scholar 

  26. de Moraes FP, Lourenco SV, Ianez RC, de Sousa EA, Silva MM, Damascena AS, Kowalski LP, Soares FA, Coutinho-Camillo CM (2017) Expression of stem cell markers in oral cavity and oropharynx squamous cell carcinoma. Oral Surg Oral Med Oral Pathol Oral Radiol 123(1):113–122. https://doi.org/10.1016/j.oooo.2016.09.009

    Article  PubMed  Google Scholar 

  27. Ohta M, Abe A, Ohno F, Hasegawa Y, Tanaka H, Maseki S, Kondo E, Kurita K, Nakanishi H (2013) Positive and negative regulation of podoplanin expression by TGF-beta and histone deacetylase inhibitors in oral and pharyngeal squamous cell carcinoma cell lines. Oral Oncol 49(1):20–26. https://doi.org/10.1016/j.oraloncology.2012.06.017S1368-8375(12)00215-1

    Article  CAS  PubMed  Google Scholar 

  28. Parhar S, Kaur H, Vashist A, Verma S (2015) Role of podoplanin in potentially malignant disorders and oral squamous cell carcinoma and its correlation with lymphangiogenesis. Indian J Cancer 52(4):617–622. https://doi.org/10.4103/0019-509X.178427 IndianJournalofCancer_2015_52_4_617_178427

    Article  CAS  PubMed  Google Scholar 

  29. Monteiro LS, Delgado ML, Ricardo S, do Amaral B, Salazar F, Pacheco JJ, Lopes CA, Bousbaa H, Warnakulasuryia S (2016) Prognostic significance of CD44v6, p63, podoplanin and MMP-9 in oral squamous cell carcinomas. Oral Dis 22(4):303–312. https://doi.org/10.1111/odi.12442

    Article  CAS  PubMed  Google Scholar 

  30. Shimada Y, Ishii G, Nagai K, Atsumi N, Fujii S, Yamada A, Yamane Y, Hishida T, Nishimura M, Yoshida J, Ikeda N, Ochiai A (2009) Expression of podoplanin, CD44, and p63 in squamous cell carcinoma of the lung. Cancer Sci 100(11):2054–2059. https://doi.org/10.1111/j.1349-7006.2009.01295.xCAS1295

    Article  CAS  PubMed  Google Scholar 

  31. Yuan P, Temam S, El-Naggar A, Zhou X, Liu DD, Lee JJ, Mao L (2006) Overexpression of podoplanin in oral cancer and its association with poor clinical outcome. Cancer 107(3):563–569. https://doi.org/10.1002/cncr.22061

    Article  CAS  PubMed  Google Scholar 

  32. Cueni LN, Hegyi I, Shin JW, Albinger-Hegyi A, Gruber S, Kunstfeld R, Moch H, Detmar M (2010) Tumor lymphangiogenesis and metastasis to lymph nodes induced by cancer cell expression of podoplanin. Am J Pathol 177(2):1004–1016. https://doi.org/10.2353/ajpath.2010.090703S0002-9440(10)60155-1

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Kreppel M, Scheer M, Drebber U, Ritter L, Zoller JE (2010) Impact of podoplanin expression in oral squamous cell carcinoma: clinical and histopathologic correlations. Virchows Arch 456(5):473–482. https://doi.org/10.1007/s00428-010-0915-7

    Article  CAS  PubMed  Google Scholar 

  34. Huber GF, Fritzsche FR, Zullig L, Storz M, Graf N, Haerle SK, Jochum W, Stoeckli SJ, Moch H (2011) Podoplanin expression correlates with sentinel lymph node metastasis in early squamous cell carcinomas of the oral cavity and oropharynx. Int J Cancer 129(6):1404–1409. https://doi.org/10.1002/ijc.25795

    Article  CAS  PubMed  Google Scholar 

  35. Garcia-Montesinos-Perea B, Val-Bernal JF, Saiz-Bustillo R (2005) Epidermoid carcinoma of the lip: an immunohistochemical study. Med Oral Patol Oral Cir Bucal 10(5):454–461

    PubMed  Google Scholar 

  36. Cruz MC, Pereira AL, Lopes FF, Nonaka CF, Silva RR, Freitas Rde A, Souza LB, Pinto LP (2009) Immunohistochemical expression of E-cadherin and CD44v6 in squamous cell carcinomas of the lower lip and tongue. Braz Dent J 20(1):64–69

    Article  PubMed  Google Scholar 

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Contributions

Alexandre Simões Garcia contributed to the study concept, design, data collection, histopathological analysis, and writing. Agnes Assao contributed to the study concept, data collection, and writing. André Lopes Carvalho contributed to the statistical analysis. Fernando Augusto Soares contributed to the study concept and providing the infra-structure. Luis Paulo Kowalski contributed to the study concept. Denise Tostes Oliveira contributed to the study concept and design and critically revised the manuscript. All authors approved the publication.

Funding

This study was funded by São Paulo Research Foundation (FAPESP)—grant #2012/13411-6 and partially supported by “Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil- CAPES”, financial code #001.

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Authors and Affiliations

  1. Department of Surgery, Stomatology, Pathology and Radiology, Area of Pathology, Bauru School of Dentistry, University of São Paulo, Alameda Octávio Pinheiro Brisola, 9-75, Bauru, SP, 17012-901, Brazil

    Alexandre Simões Garcia, Agnes Assao & Denise Tostes Oliveira

  2. Fundação Pio XII Institution – Cancer Hospital of Barretos, Brazil, R. Antenor Duarte Viléla, 1331, Barretos, SP, 14784-400, Brazil

    André Lopes Carvalho

  3. Rede D’Or Hospitals Network – Pathology Division – Brazil, R. das Perobas, 344, Jabaquara, São Paulo, SP, 04321-120, Brazil

    Fernando Augusto Soares

  4. Department of Head and Neck Surgery and Otorhinolaryngology, A.C. Camargo Cancer Hospital, R. Professor Antônio Prudente, 211, Liberdade, São Paulo, SP, 01509-010, Brazil

    Luiz Paulo Kowalski

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  1. Alexandre Simões Garcia
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  2. Agnes Assao
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Correspondence to Denise Tostes Oliveira.

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The Research Ethics Committee of the A.C. Camargo Cancer Center, São Paulo, Brazil, approved this study (process #1778/13 - CAAE 01594912.0.3001.5432).

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Garcia, A.S., Assao, A., Carvalho, A.L. et al. The stem cell markers expression CD44v6 and podoplanin in lip cancer: clinical significance. Virchows Arch 474, 745–754 (2019). https://doi.org/10.1007/s00428-019-02539-3

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