Histochemistry and Cell Biology

, Volume 131, Issue 5, pp 651–660 | Cite as

Expression of Snail is associated with myofibroblast phenotype development in oral squamous cell carcinoma

  • Marcus Franz
  • Karin Spiegel
  • Claudia Umbreit
  • Petra Richter
  • Carolina Codina-Canet
  • Angela Berndt
  • Annelore Altendorf-Hofmann
  • Sven Koscielny
  • Peter Hyckel
  • Hartwig Kosmehl
  • Ismo Virtanen
  • Alexander Berndt
Original Paper
First Online:
Accepted:

Abstract

Snail is a regulator of epithelial–mesenchymal transition (EMT) and considered crucial to carcinoma metastasis, myofibroblast transdifferentiation, and fibroblast activation. To investigate the role of Snail in oral squamous cell carcinoma (OSCC), its immunohistochemical expression was analysed in 129 OSCC samples and correlated to nodal metastasis, histological grade, E-cadherin, and alpha smooth-muscle-actin (αSMA). The results were compared to findings in 23 basal cell carcinomas (BCC). Additionally, the influence of TGFβ1 and EGF on Snail, E-cadherin, vimentin, and αSMA expression was analysed in two OSCC cell lines. As a result, Snail-positive cells were mainly found in the stroma of the OSCC invasive front without statistically significant correlation to histological grade or nodal metastasis. Snail was co-localised to αSMA but not to E-cadherin or cytokeratin and showed a significant correlation to the loss of membranous E-cadherin. All BCCs were Snail negative. In OSCC culture, the growth-factor-mediated EMT-like phenomenon was accompanied by αSMA down-regulation. In summary, Snail expression in OSCC is a stromal phenomenon associated with the myofibroblast phenotype and not related to growth-factor-mediated transdifferentiation of the carcinoma cells themselves. Consequently, Snail immunohistochemistry cannot contribute to the prediction of the metastatic potential. Furthermore, stromal Snail expression is suggested to be the result of mutual paracrine interaction of fibro-/myofibroblasts and dedifferentiated carcinoma cells leading to the generation of a special type of carcinoma-associated fibroblasts.

Keywords

Snail Oral squamous cell carcinoma Metastasis Myofibroblast Alpha smooth-muscle-actin 
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Notes

Acknowledgements

The authors would like to thank Susanne Bergmann, Christiane Geier, Angela Gröbner, and Katrin Schlehahn for excellent technical assistance. The study was supported by the Thuringian Ministry of Science, Research and Art (ThMWFK), and IZKF of the University Hospital Jena.

References

  1. Barnes L, Eveson JW, Reichart PA, Sidransky D (2005) Pathology and genetics of head and neck tumours (World Health Organization Classification of Tumours). IARC Press, LyonGoogle Scholar
  2. Barth PJ, Ebrahimsade S, Ramaswamy A, Moll R (2002) CD34+ fibrocytes in invasive ductal carcinomas, ductal carcinoma in situ, and benign breast lesions. Virchows Arch 440:298–303PubMedCrossRefGoogle Scholar
  3. Barth PJ, Schenk zu Schweinsberg T, Ramaswamy A, Moll R (2004) CD34+ fibrocytes, α-smooth muscle antigen-positive myofibroblasts, and CD117 expression in the stroma of invasive squamous cell carcinomas of the oral cavity, pharynx, and larynx. Virchows Arch 444:231–234PubMedCrossRefGoogle Scholar
  4. Batlle E, Sancho E, Franci C, Dominguez D, Monfar M, Baulida J, Garcia De Hereros A (2000) The transcription factor Snail is a repressor of E-cadherin gene expression in epithelial tumour cells. Nat Cell Biol 2:84–89PubMedCrossRefGoogle Scholar
  5. Berndt A, Hyckel P, Könneker A, Katenkamp D, Kosmehl H (1997) Oral squamous cell carcinoma invasion is associated with a laminin-5 matrix re-organization but independent of basement membrane hemidesmosome formation. Clues from an in vitro model. Invasion Metastasis 17:251–258PubMedGoogle Scholar
  6. Berndt A, Borsi L, Hyckel P, Kosmehl H (2001) Fibrillary co-deposition of laminin-5 and large unspliced tenascin-C in the invasive front of oral squamous cell carcinoma in vivo and in vitro. J Cancer Res Clin Oncol 127:286–292PubMedCrossRefGoogle Scholar
  7. Blanco MJ, Moreno-Bueno G, Sartio D, Locasicio A, Cano A, Palacios J, Nieto MA (2002) Correlation of Snail expression with histological grade and lymph node status in breast carcinomas. Oncogene 21:3241–3246PubMedCrossRefGoogle Scholar
  8. Bolos V, Peinado H, Perez-Moreno MA, Fraga MF, Esteller M, Cano A (2003) The transcription factor Slug represses E-cadherin expression and induces epithelial to mesenchymal transitions: a comparison with Snail and E47 repressors. J Cell Sci 116:499–511PubMedCrossRefGoogle Scholar
  9. Boutet A, Esteban MA, Maxwell PH, Nieto MA (2007) Reactivation of Snail genes in renal fibrosis and carcinomas. A process of reversed embryogenesis? Cell Cycle 6:638–642PubMedGoogle Scholar
  10. Bryne M, Koppang HS, Lilleng R, Kjaerheim A (1992) Malignancy grading of the deep invasive margins of oral squamous cell carcinomas has high prognostic value. J Pathol 166:375–381PubMedCrossRefGoogle Scholar
  11. Cano A, Perez-Moreno MA, Rodrigo I, Locascio A, Blanco MJ, del Barrio MG, Portillo F, Nieto MA (2000) The transcription factor Snail controls epithelial–mesenchymal transitions by repressing E-cadherin expression. Nat Cell Biol 2:76–83PubMedCrossRefGoogle Scholar
  12. Daly AJ, McIlreavey L, Irwin CR (2008) Regulation of HGF and SDF-1 expression by oral fibroblasts—implications for invasion of oral cancer. Oral Oncol 44:646–651PubMedCrossRefGoogle Scholar
  13. De Rosa G, Barra E, Guarino M, Staibano S, Donofrio V, Boscaino A (1994) Fibronectin, laminin, type IV collagen distribution, and myofibroblastic stromal reaction in aggressive and nonaggressive basal cell carcinoma. Am J Dermatopathol 16:258–267PubMedCrossRefGoogle Scholar
  14. De Vries AS, Tilton RG, Mortier S, Lameire NH (2006) Myofibroblast transdifferentiation of mesothelial cells is mediated by RAGE and contributes to peritoneal fibrosis in uraemia. Nephrol Dial Transplant 21:2549–2555CrossRefGoogle Scholar
  15. De Wever O, Mareel M (2003) Role of tissue stroma in cancer cell invasion. J Pathol 200:429–447PubMedCrossRefGoogle Scholar
  16. Franci C, Takkunen M, Dave N, Alameda F, Gómez S, Rodríguez R, Escrivà M, Montserrat-Sentís B, Baró T, Garrido M, Bonilla F, Virtanen I, García de Herreros A (2006) Expression of Snail protein in tumor–stroma interface. Oncogene 25(37):5134–5144PubMedGoogle Scholar
  17. Franz M, Hansen T, Richter P, Borsi L, Böhmer FD, Hyckel P, Schleier P, Katenkamp D, Zardi L, Kosmehl H, Berndt A (2006) Complex formation of the laminin-5 gamma2 chain and large unspliced tenascin-C in oral squamous cell carcinoma in vitro and in situ: implications for sequential modulation of extracellular matrix in the invasive tumor front. Histochem Cell Biol 126(1):125–131PubMedCrossRefGoogle Scholar
  18. Franz M, Richter P, Geyer C, Hansen T, Acuna LD, Hyckel P, Bohmer FD, Kosmehl H, Berndt A (2007) Mesenchymal cells contribute to the synthesis and deposition of the laminin-5 gamma 2 chain in the invasive front of oral squamous cell carcinoma. J Mol Histol 38(3):183–190PubMedCrossRefGoogle Scholar
  19. Giannelli G, Bergamini C, Fransvea E, Sqarra C, Antonaci S (2005) Laminin-5 with transforming growth factor-beta1 induces epithelial to mesenchymal transition in hepatocellular carcinoma. Gastroenterology 129:1375–1383PubMedCrossRefGoogle Scholar
  20. Guaita S, Puig I, Franci C, Garrido M, Dominguez D, Batlle E, Sancho E, Dedhar S, De Herreros AG, Baulida J (2002) Snail induction of epithelial to mesenchymal transition in tumor cells is accompanied by MUC1 repression and ZEB1 expression. J Biol Chem 277:39209–39216PubMedCrossRefGoogle Scholar
  21. Higashikawa K, Yoneda S, Taki M, Shigeishi H, Ono S, Tobiume K, Kamata N (2008) Gene expression profiling to identify genes associated with high-invasiveness in human squamous cell carcinoma with epithelial-to-mesenchymal transition. Cancer Lett 264:256–264PubMedCrossRefGoogle Scholar
  22. Hung SC, Kuo PY, Chang CF, Chen TH, Ho LL (2006) Alpha-smooth muscle actin expression and structure integrity in chondrogenesis of human mesenchymal stem cells. Cell Tissue Res 324:457–466PubMedCrossRefGoogle Scholar
  23. Iwano M, Plieth D, Danoff TM, Xue C, Okada H, Neilson EG (2002) Evidence that fibroblasts derive from epithelium during tissue fibrosis. J Clin Invest 110:341–350PubMedGoogle Scholar
  24. Jin X, Li J, Li Z, Li Y (2001) Expression of transforming growth factor beta (TGF-beta) subtypes in oral squamous cell carcinoma. Hua Xi Kou Qiang Yi Xue Za Zhi 19:377–379PubMedGoogle Scholar
  25. Kalluri R, Neilson EG (2003) Epithelial–mesenchymal transition and its implication for fibrosis. J Clin Invest 112:1776–1784PubMedGoogle Scholar
  26. Kellermann MG, Sobral LM, da Silva SD, Zecchin KG, Graner E, Lopes MA, Kowalski LP, Coletta RD (2008) Mutual paracrine effects of oral squamous cell carcinoma cells and normal oral fibroblasts: induction of fibroblast to myofibroblast transdifferentiation and modulation of tumor cell proliferation. Oral Oncol 44:509–517PubMedCrossRefGoogle Scholar
  27. Lee JM, Dedhar S, Kalluri R, Thompson EW (2006) The epithelial-mesenchymal transition: new insights in signalling, development, and disease. J Cell Biol 172:973–981PubMedCrossRefGoogle Scholar
  28. Lewis MP, Lygoe KA, Nystrom ML, Anderson WP, Speight PM, Marshall JF, Thomas GJ (2004) Tumour-derived TGF-beta1 modulates myofibroblast differentiation and promotes HGF/SF-dependent invasion of squamous carcinoma cells. Br J Cancer 90(4):822–832PubMedCrossRefGoogle Scholar
  29. Maeda G, Chiba T, Okazaki M, Satoh T, Taya Y, Aoba T, Kato K, Kawashiri S, Imai K (2005) Expression of SIP1 in oral squamous cell carcinomas: implications for E-cadherin expression and tumour progression. Int J Oncol 27:1535–1541PubMedGoogle Scholar
  30. Meindl-Beinker NM, Dooley S (2008) Transforming growth factor-beta and hepatocyte transdifferentiation in liver fibrosis. J Gastroenterol Hepatol 23(Suppl 1):S122–S127PubMedCrossRefGoogle Scholar
  31. Natsugoe S, Uchikado Y, Okumura H, Matsumoto M, Setoyama T, Tamotsu K, Kita Y, Sakamoto A, Owaki T, Ishigami S, Aikou T (2007) Snail plays a key role in E-cadherin-preserved esophageal squamous cell carcinoma. Oncol Rep 17:517–523PubMedGoogle Scholar
  32. Onoue T, Uchida D, Bequm NM, Tomizuka Y, Yoshida H, Sato M (2006) Epithelial-mesenchymal transition induced by the stromal cell-derived factor-1/CXCR4 system in oral squamous cell carcinoma cells. Int J Oncol 29:1133–1138PubMedGoogle Scholar
  33. Peiro S, Escriva M, Puig I, Barberà MJ, Dave N, Herranz N, Larriba MJ, Takkunen M, Francí C, Muñoz A, Virtanen I, Baulida J, García de Herreros A (2006) Snail 1 transcriptional repressor binds to its own promoter and controls its expression. Nucleic Acids Res 34:2077–2084PubMedCrossRefGoogle Scholar
  34. Pena C, Garcia JM, Silva J, García V, Rodríguez R, Alonso I, Millán I, Salas C, de Herreros AG, Muñoz A, Bonilla F (2005) E-cadherin and vitamin D receptor regulation by Snail and Zeb1 in colon cancer: clinicopathological correlations. Hum Mol Genet 14:3361–3370PubMedCrossRefGoogle Scholar
  35. Petersen OW, Lind Nielsen H, Gudjonsson T, Villadseu R, Rønnov-Jessen L, Bissell ML (2001) The plasticity of human breast carcinoma cells is more than epithelial to mesenchymal conversion. Breast Cancer Res 3(4):213–217PubMedCrossRefGoogle Scholar
  36. Petersen OW, Nielsen H, Gudjonsson T, Villadsen R, Rank F, Niebuhr E, Bissell MJ, Rønnov-Jessen L (2003) Epithelial to mesenchymal transition in human breast cancer can provide a nonmalignant stroma. Am J Pathol 162:391–402PubMedGoogle Scholar
  37. Prindull G, Zipori D (2004) Environmental guidance of normal and tumor cell plasticity: epithelial mesenchymal transitions as a paradigm. Blood 103:2892–2899PubMedCrossRefGoogle Scholar
  38. Radisky DC, Kenny PA, Bissell MJ (2007) Fibrosis and cancer: do myofibroblasts come also from epithelial cells via EMT? J Cell Biochem 101(4):830–839PubMedCrossRefGoogle Scholar
  39. Ronnov-Jessen L, Petersen OW, Koteliansky VE, Bissell MJ (1995) The origin of the myofibroblast in breast cancer: recapitulation of tumor environment in culture unreveals diversity and implicates converted fibroblasts and recruited smooth muscle cells. J Clin Invest 95:859–873PubMedCrossRefGoogle Scholar
  40. Stuelten CH, DaCosta Byfield S, Arany PR, Karpova TS, Stetler-Stevenson WG, Roberts AB (2005) Breast cancer cells induce stromal fibroblasts to express MMP-9 via secretion of TNF-alpha and TGF-beta. J Cell Sci 118:2143–2153PubMedCrossRefGoogle Scholar
  41. Sun L, Diamond ME, Ottaviano AJ, Joseph MJ, Ananthanarayan V, Munshi HG (2008) Transforming growth factor-beta 1 promotes matrix metalloproteinase-9-mediated oral cancer invasion through Snail expression. Mol Cancer Res 6:10–20PubMedCrossRefGoogle Scholar
  42. Taki M, Higashikawa K, Yoneda S, Ono S, Shigeishi H, Nagayama M, Kamata N (2008) Up-regulation of stromal cell-derived factor-1alpha and its receptor CXCR4 expression accompanied with epithelial–mesenchymal transition in human oral squamous cell carcinoma. Oncol Rep 19:993–998PubMedGoogle Scholar
  43. Takkunen M, Grenman R, Hukkanen M, Korhonen M, Garcia de Herreros A, Virtanen I (2006) Snail-dependent and -independent epithelial–mesenchymal transition in oral squamous carcinoma cells. J Histochem Cytochem 54(11):1263–1275PubMedCrossRefGoogle Scholar
  44. Takkunen M, Ainola M, Vainionpää N, Grenman R, Patarroyo M, Garcia de Herreros A, Konttinen YT, Virtanen I (2008) Epithelial–mesenchymal transition downregulates laminin a5 chain and upregulates a4 chain in oral squamous carcinoma cells. Histochem Cell Biol 130:509–525PubMedCrossRefGoogle Scholar
  45. Thiery JP (2002) Epithelial–mesenchymal transition in tumour progression. Nat Rev Cancer 2:442–454PubMedCrossRefGoogle Scholar
  46. Thompson LDR, Wieneke JA, Miettinen M, Heffner DK (2002) Spindle cell (sarcomatoid) carcinoma of the larynx. A clinicopathologic study of 187 cases. Am J Surg Pathol 26:153–170PubMedCrossRefGoogle Scholar
  47. Wilkins-Port CE, Higgins PJ (2007) Regulation of extracellular matrix remodelling following transforming growth factor-β1/epidermal growth factor-stimulated epithelial–mesenchymal transition in human premalignant keratinocytes. Cells Tissues Organs 185:116–122PubMedCrossRefGoogle Scholar
  48. Yang J, Mani SA, Donaher JL, Ramaswamy S, Itzykson RA, Come C, Savagner P, Gitelman I, Richardson A, Weinberg RA (2004) Twist, a master regulator of morphogenesis, plays an essential role in tumor metastasis. Cell 117:927–939PubMedCrossRefGoogle Scholar
  49. Yanjia H, Xinchun J (2007) The role of epithelial-mesenchymal transition in oral squamous cell carcinoma and oral submucous fibrosis. Clin Chim Acta 383:51–56PubMedCrossRefGoogle Scholar
  50. Yin T, Wang CY, Liu T, Thao G, Zha YH (2006) Expression of Snail and E-cadherin in pancreatic carcinoma and clinical significance thereof. Zhonghua Yi Xue Za Zhi 86:2821–2925PubMedGoogle Scholar
  51. Yokoyama K, Kamata N, Fujimoto R, Tsutsumi S, Tomonari M, Taki M, Hosokawa H, Nagayama M (2003) Increased invasion and matrix metalloproteinase-2 expression by Snail-induced mesenchymal transition in squamous cell carcinoma. Int J Oncol 22:891–898PubMedGoogle Scholar
  52. Zhang W, Alt-Holland A, Margulis A, Shamis Y, Fusenig NE, Rodeck U, Garlick JA (2006) E-cadherin loss promotes the initiation of squamous cell carcinoma invasion through modulation of integrin-mediated adhesion. J Cell Sci 119:283–291PubMedCrossRefGoogle Scholar
  53. Zidar N, Gale N, Kojc N, Volavsek M, Cardesa A, Alos L, Höfler H, Blechschmidt K, Becker KF (2008) Cadherin-catenin complex and transcription factor Snail-1 in spindle cell carcinoma of the head and neck. Virchows Arch 453:267–274PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Marcus Franz
    • 1
  • Karin Spiegel
    • 5
  • Claudia Umbreit
    • 2
  • Petra Richter
    • 2
  • Carolina Codina-Canet
    • 2
  • Angela Berndt
    • 3
  • Annelore Altendorf-Hofmann
    • 4
  • Sven Koscielny
    • 5
  • Peter Hyckel
    • 6
  • Hartwig Kosmehl
    • 7
  • Ismo Virtanen
    • 8
  • Alexander Berndt
    • 2
  1. 1.Clinic of Internal Medicine IUniversity Hospital JenaJenaGermany
  2. 2.Institute of PathologyUniversity Hospital JenaJenaGermany
  3. 3.Friedrich-Loeffler-InstituteInstitute of Molecular PathogenesisJenaGermany
  4. 4.Tumour RegistraryUniversity Hospital JenaJenaGermany
  5. 5.ENT DepartmentUniversity Hospital JenaJenaGermany
  6. 6.Clinic of Maxillofacial Surgery/Plastic SurgeryUniversity Hospital JenaJenaGermany
  7. 7.Institute of PathologyHELIOS-KlinikumErfurtGermany
  8. 8.Institute of Biomedicine/AnatomyUniversity of HelsinkiHelsinkiFinland

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