Virchows Archiv

, Volume 466, Issue 5, pp 559–569 | Cite as

Keratin 17 is co-expressed with 14-3-3 sigma in oral carcinoma in situ and squamous cell carcinoma and modulates cell proliferation and size but not cell migration

  • Toshihiko Mikami
  • Satoshi Maruyama
  • Tatsuya Abé
  • Takanori Kobayashi
  • Manabu Yamazaki
  • Akinori Funayama
  • Susumu Shingaki
  • Tadaharu Kobayashi
  • Cheng Jun
  • Takashi SakuEmail author
Original Article


Expression of keratin (K) 13 is replaced with that of K17 when squamous cells of the oral mucosa transform from normal and dysplastic epithelia to carcinoma in situ (CIS) and squamous cell carcinoma (SCC). Since 14-3-3 sigma is functionally associated with K17, we examined possible relationships between expression of K17 and 14-3-3 sigma in oral CIS and SCC tissues by immunohistochemistry. We furthermore examined whether or not K17 expression or knockdown by small interfering RNA (siRNA) modulates the behavior of SCC cells in culture in terms of cell proliferation and migration. In tissue specimens of oral SCC and CIS, the pattern of cytoplasmic expression of 14-3-3 sigma and K17 was similar but neither was expressed in normal or dysplastic epithelia. Both proteins were demonstrated in the cytoplasm of control oral SCC ZK-1 cells, but expression of 14-3-3 sigma changed from cytoplasmic to nuclear upon knockdown of K17. In carcinoma cells, therefore, cytoplasmic localization of 14-3-3 sigma seems to accompany expression of K17. In K17-knockdown cells, proliferation was significantly suppressed at 4 days after seeding. In addition, the cell size of K17-knockdown cells was significantly smaller than that of control cells; as a result of which in the migration experiments, we found delayed closure of scratch wounds but migration as such was not affected. We conclude that K17 expression promotes SCC cell growth and cell size but does not affect cell migration. K17 expression is accompanied by cytoplasmic expression of 14-3-3 sigma, indicative of their functional relationship.


Oral squamous cell carcinoma Carcinoma in situ Keratin 17 14-3-3 sigma siRNA 



This work was supported in part by Grants-in-Aid for Scientific Research (to SM, JC, and TS) and for Young Scientists (to TM and SM) from the Japan Society for the Promotion of Science and by a grant for the Promotion of Niigata University Research Projects (to TM and SM).

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

428_2015_1735_MOESM1_ESM.pdf (182 kb)
Online Resource 1 Unusual case of moderate epithelial dysplasia case positive for 14-3-3 sigma. (a) H&E stain, immunoperoxidase stains for 14-3-3 sigma (b), K17 (c), K13 (d), and Ki-67 (e), hematoxylin counterstain. a: × 200; b-e: × 300. In five cases of epithelial dysplasia (a), 14-3-3 sigma was weakly positive in the whole epithelial layer (b), while no staining for K17 (c) was noted. K13 was occasionally positive in the prickle cell layer (d), and Ki-67-positive cells were sporadically detected only in the second basal layer (e). This pattern of marker expression is characteristic for moderate dysplasia. (PDF 177 kb)


  1. 1.
    Kobayashi T, Maruyama S, Cheng J, Ida-Yonemochi H, Yagi M, Takagi R, Saku T (2010) Histopathological varieties of oral carcinoma in situ: diagnosis aided by immunohistochemistry dealing with the second basal cell layer as the proliferating center of oral mucosal epithelia. Pathol Int 60:156–166CrossRefPubMedGoogle Scholar
  2. 2.
    Funayama A, Cheng J, Maruyama S, Yamazaki M, Kobayashi T, Syafriadi M, Kundu S, Shingaki S, Saito C, Saku T (2011) Enhanced expression of podoplanin in oral carcinomas in situ and squamous cell carcinomas. Pathobiology 78:171–180CrossRefPubMedGoogle Scholar
  3. 3.
    Mikami T, Cheng J, Maruyama S, Kobayashi T, Funayama A, Yamazaki M, Adeola HA, Wu L, Shingaki S, Saito C, Saku T (2011) Emergence of keratin 17 vs. loss of keratin 13: their reciprocal immunohistochemical profiles in oral carcinoma in situ. Oral Oncol 47:497–503CrossRefPubMedGoogle Scholar
  4. 4.
    Kobayashi T, Maruyama S, Abé T, Cheng J, Takagi R, Saito C, Saku T (2012) Keratin-10-positive orthokeratotic dysplasia: a new leukoplakia-type precancerous entity of the oral mucosa. Histopathology 61:910–920CrossRefPubMedGoogle Scholar
  5. 5.
    Funayama A, Maruyama S, Yamazaki M, Al-Eryani K, Shingaki S, Saito C, Cheng J, Saku T (2012) Intraepithelially entrapped blood vessels in oral carcinoma in-situ. Virchows Arch 460:473–480CrossRefPubMedGoogle Scholar
  6. 6.
    Ida-Yonemochi H, Maruyama S, Kobayashi T, Yamazaki M, Cheng J, Saku T (2012) Loss of keratin 13 in oral carcinoma in situ: a comparative study of protein and gene expression levels using paraffin sections. Mod Pathol 25:784–794CrossRefPubMedGoogle Scholar
  7. 7.
    Ikarashi T, Ida-Yonemochi H, Ohshiro K, Cheng J, Saku T (2004) Intraepithelial expression of perlecan, a basement membrane-type heparan sulfate proteoglycan reflects dysplastic changes of the oral mucosal epithelium. J Oral Pathol Med 33:87–95CrossRefPubMedGoogle Scholar
  8. 8.
    Tilakaratne WM, Kobayashi T, Ida-Yonemochi H, Swelam W, Yamazaki M, Mikami T, Alvarado CG, Shahidul AM, Maruyama S, Cheng J, Saku T (2009) Matrix metalloproteinase 7 and perlecan in oral epithelial dysplasia and carcinoma in situ: an aid for histopathologic recognition of their cell proliferation centers. J Oral Pathol Med 38:348–355CrossRefPubMedGoogle Scholar
  9. 9.
    Metwaly H, Maruyama S, Yamazaki M, Tsuneki M, Abé T, Jen KY, Cheng J, Saku T (2012) Parenchymal-stromal switching for extracellular matrix production on invasion of oral squamous cell carcinoma. Hum Pathol 43:1973–1981CrossRefPubMedGoogle Scholar
  10. 10.
    Alvarado CG, Maruyama S, Cheng J, Ida-Yonemochi H, Kobayashi T, Yamazaki M, Takagi R, Saku T (2011) Nuclear translocation of beta-catenin synchronized with loss of E-cadherin in oral epithelial dysplasia with a characteristic two-phase appearance. Histopathology 59:283–291PubMedGoogle Scholar
  11. 11.
    Troyanovsky SM, Guelstein VI, Tchipysheva TA, Krutovskikh VA, Bannikov GA (1989) Patterns of expression of keratin 17 in human epithelia: dependency on cell position. J Cell Sci 93:419–426PubMedGoogle Scholar
  12. 12.
    Kim S, Wong P, Coulombe PA (2006) A keratin cytoskeletal protein regulates protein synthesis and epithelial cell growth. Nature 441:362–365CrossRefPubMedGoogle Scholar
  13. 13.
    Maruyama S, Cheng J, Yamazaki M, Zhou XJ, Zhang ZY, He RG, Saku T (2010) Metastasis-associated genes in salivary adenoid cystic carcinoma and oral squamous cell carcinoma: a differential DNA chip analysis between metastatic and non-metastatic cell systems. Cancer Genet Cytogenet 195:14–22CrossRefGoogle Scholar
  14. 14.
    Syafriadi M, Cheng J, Jen KY, Ida-Yonemochi H, Suzuki M, Saku T (2005) Two-phase appearance of oral epithelial dysplasia resulting from focal proliferation of parabasal cells and apoptosis of prickle cells. J Oral Pathol Med 34:140–149CrossRefPubMedGoogle Scholar
  15. 15.
    Smith FJ, Jonkman MF, van Goor H, Coleman CM, Covello SP, Uitto J, McLean WH (1998) A mutation in human keratin K6b produces a phenocopy of the K17 disorder pachyonychia congenita type 2. Hum Mol Genet 7:1143–1148CrossRefPubMedGoogle Scholar
  16. 16.
    Troyanovsky SM, Leube RE, Franke WW (1992) Characterization of the human gene encoding cytokeratin 17 and its expression pattern. Eur J Cell Biol 59:127–137PubMedGoogle Scholar
  17. 17.
    McGowan KM, Coulombe PA (1998) Onset of keratin 17 expression coincides with the definition of major epithelial lineages during skin development. J Cell Biol 143:469–486CrossRefPubMedCentralPubMedGoogle Scholar
  18. 18.
    Freedberg IM, Tomic-Canic M, Komine M, Blumenberg M (2001) Keratins and the keratinocyte activation cycle. J Investig Dermatol 116:633–640CrossRefPubMedGoogle Scholar
  19. 19.
    Kozma SC, Thomas G (2002) Regulation of cell size in growth, development and human disease: PI3K, PKB and S6K. Bioessays 24:65–71CrossRefPubMedGoogle Scholar
  20. 20.
    Bertram PG, Zeng C, Thorson J, Shaw AS, Zheng XF (1998) The 14-3-3 proteins positively regulate rapamycin-sensitive signaling. Curr Biol 8:1259–1267CrossRefPubMedGoogle Scholar
  21. 21.
    Yang HY, Wen YY, Chen CH, Lozano G, Lee MH (2003) 14-3-3 sigma positively regulates p53 and suppresses tumor growth. Mol Cell Biol 23:7096–7107CrossRefPubMedCentralPubMedGoogle Scholar
  22. 22.
    Chan TA, Hwang PM, Hermeking H, Kinzler KW, Vogelstein B (2000) Cooperative effects of genes controlling the G(2)/M checkpoint. Genes Dev 14:1584–1588PubMedCentralPubMedGoogle Scholar
  23. 23.
    Strimpakos AS, Karapanagiotou EM, Saif MW, Syrigos KN (2009) The role of mTOR in the management of solid tumors: an overview. Cancer Treat Rev 35:148–159CrossRefPubMedGoogle Scholar
  24. 24.
    Janus A, Robak T, Smolewski P (2005) The mammalian target of the rapamycin (mTOR) kinase pathway: its role in tumourigenesis and targeted antitumour therapy. Cell Mol Biol Lett 10:479–498PubMedGoogle Scholar
  25. 25.
    Asnaghi L, Bruno P, Priulla M, Nicolin A (2004) mTOR: a protein kinase switching between life and death. Pharmacol Res 50:545–549CrossRefPubMedGoogle Scholar
  26. 26.
    Xu G, Zhang W, Bertram P, Zheng XF, McLeod H (2004) Pharmacogenomic profiling of the PI3K/PTEN-AKT-mTOR pathway in common human tumors. Int J Oncol 24:893–900PubMedGoogle Scholar
  27. 27.
    Hay N, Sonenberg N (2004) Upstream and downstream of mTOR. Genes Dev 18:1926–1945CrossRefPubMedGoogle Scholar
  28. 28.
    Hayashi E, Kuramitsu Y, Fujimoto M, Zhang X, Tanaka T, Uchida K, Fukuda T, Furumoto H, Ueyama Y, Nakamura K (2009) Proteomic profiling of differential display analysis for human oral squamous cell carcinoma: 14-3-3 σ protein is upregulated in human oral squamous cell carcinoma and dependent on the differentiation level. Proteomics Clin Appl 3:1338–1347CrossRefPubMedGoogle Scholar
  29. 29.
    Kirschner M, Montazem A, Hilaire HS, Radu A (2006) Long-term culture of human gingival keratinocyte progenitor cells by down-regulation of 14-3-3 sigma. Stem Cells Dev 15:556–565CrossRefPubMedGoogle Scholar
  30. 30.
    Takahashi Y, Nishikawa M, Takakura Y (2006) Suppression of tumor growth by intratumoral injection of short hairpin RNA-expressing plasmid DNA targeting beta-catenin or hypoxia-inducible factor 1alpha. J Control Release 116:90–95CrossRefPubMedGoogle Scholar
  31. 31.
    Wang Z, Rao DD, Senzer N, Nemunaitis J (2011) RNA interference and cancer therapy. Pharm Res 28:2983–2995CrossRefPubMedGoogle Scholar
  32. 32.
    Liu X, Sempere LF, Ouyang H, Memoli VA, Andrew AS, Luo Y, Demidenko E, Korc M, Shi W, Preis M, Dragnev KH, Li H, Direnzo J, Bak M, Freemantle SJ, Kauppinen S, Dmitrovsky E (2010) MicroRNA-31 functions as an oncogenic microRNA in mouse and human lung cancer cells by repressing specific tumor suppressors. J Clin Invest 120:1298–1309CrossRefPubMedCentralPubMedGoogle Scholar
  33. 33.
    Santel A, Aleku M, Röder N, Möpert K, Durieux B, Janke O, Keil O, Endruschat J, Dames S, Lange C, Eisermann M, Löffler K, Fechtner M, Fisch G, Vank C, Schaeper U, Giese K, Kaufmann J (2010) Atu027 prevents pulmonary metastasis in experimental and spontaneous mouse metastasis models. Clin Cancer Res 16:5469–5480CrossRefPubMedGoogle Scholar
  34. 34.
    Davis ME, Zuckerman JE, Choi CH, Seligson D, Tolcher A, Alabi CA, Yen Y, Heidel JD, Ribas A (2010) Evidence of RNAi in humans from systemically administered siRNA via targeted nanoparticles. Nature 464:1067–1070CrossRefPubMedCentralPubMedGoogle Scholar
  35. 35.
    Patnaik A, Chiorean EG, Tolcher A, Papadopoulos K, Beeram M, Kee D, Waddell M, Gilles E, Buchbinder A (2009) EZN-2968, a novel hypoxia-inducible factor-1 alpha (HIF-1 alpha) messenger ribonucleic acid (mRNA) antagonist: results of a phase I, pharmacokinetic (PK), dose-escalation study of daily administration in patients (pts) with advanced malignancies. J Clin Oncol 27:2564Google Scholar
  36. 36.
    Senzer N, Barve M, Kuhn J, Melnyk A, Beitsch P, Lazar M, Lifshitz S, Magee M, Oh J, Mill SW, Bedell C, Higgs C, Kumar P, Yu Y, Norvell F, Phalon C, Taquet N, Rao DD, Wang Z, Jay CM, Pappen BO, Wallraven G, Brunicardi FC, Shanahan DM, Maples PB, Nemunaitis J (2012) Phase I trial of "bi-shRNAi(furin)/GMCSF DNA/autologous tumor cell" vaccine (FANG) in advanced cancer. Mol Ther 20:679–686CrossRefPubMedCentralPubMedGoogle Scholar
  37. 37.
    Wei KJ, Zhang L, Yang X, Zhong LP, Zhou XJ, Pan HY, Li J, Chen WT, Zhang ZY (2009) Overexpression of cytokeratin 17 protein in oral squamous cell carcinoma in vitro and in vivo. Oral Dis 15:111–117CrossRefPubMedGoogle Scholar
  38. 38.
    Sarbia M, Fritze F, Geddert H, von Weyhern C, Rosenberg R, Gellert K (2007) Differentiation between pancreaticobiliary and upper gastrointestinal adenocarcinomas: is analysis of cytokeratin 17 expression helpful? Am J Clin Pathol 128:255–259CrossRefPubMedGoogle Scholar
  39. 39.
    van de Rijn M, Perou CM, Tibshirani R, Haas P, Kallioniemi O, Kononen J, Torhorst J, Sauter G, Zuber M, Köchli OR, Mross F, Dieterich H, Seitz R, Ross D, Botstein D, Brown P (2002) Expression of cytokeratins 17 and 5 identifies a group of breast carcinomas with poor clinical outcome. Am J Pathol 161:1991–1996CrossRefPubMedCentralPubMedGoogle Scholar
  40. 40.
    Yoshikawa K, Katagata Y, Kondo S (1998) Biochemical and immunohistochemical analyses of keratin expression in basal cell carcinoma. J Dermatol Sci 17:15–23CrossRefPubMedGoogle Scholar
  41. 41.
    Ikeda K, Tate G, Suzuki T, Mitsuya T (2008) Coordinate expression of cytokeratin 8 and cytokeratin 17 immunohistochemical staining in cervical intraepithelial neoplasia and cervical squamous cell carcinoma: an immunohistochemical analysis and review of the literature. Gynecol Oncol 108:598–602CrossRefPubMedGoogle Scholar
  42. 42.
    Carrilho C (2004) Keratins 8, 10, 13, and 17 are useful markers in the diagnosis of human cervix carcinomas. Hum Pathol 35:546–551CrossRefPubMedGoogle Scholar
  43. 43.
    Cohen-Kerem R, Madah W, Sabo E, Rahat MA, Greenberg E, Elmalah I (2004) Cytokeratin-17 as a potential marker for squamous cell carcinoma of the larynx. Ann Otol Rhinol Laryngol 113:821–827CrossRefPubMedGoogle Scholar
  44. 44.
    Lyda MH, Weiss LM (2000) Immunoreactivity for epithelial and neuroendocrine antibodies are useful in the differential diagnosis of lung carcinomas. Hum Pathol 31:980–987CrossRefPubMedGoogle Scholar
  45. 45.
    Saito M, Kobayashi T, Takagi R, Saku T (2012) Clinicopathological distinction of two categories of oral squamous cell carcinoma of the tongue: de novo vs. sequential types. Oral Med Pathol 16:81–88CrossRefGoogle Scholar
  46. 46.
    Shegokar R, Al Shaal L, Mishra PR (2011) SiRNA delivery: challenges and role of carrier systems. Pharmazie 66:313–318PubMedGoogle Scholar
  47. 47.
    Dominska M, Dykxhoorn DM (2010) Breaking down the barriers: siRNA delivery and endosome escape. J Cell Sci 123:1183–1189CrossRefPubMedGoogle Scholar
  48. 48.
    Takanashi M, Oikawa K, Sudo K, Tanaka M, Fujita K, Ishikawa A, Nakae S, Kaspar RL, Matsuzaki M, Kudo M, Kuroda M (2009) Therapeutic silencing of an endogenous gene by siRNA cream in an arthritis model mouse. Gene Ther 16:982–989CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Toshihiko Mikami
    • 1
    • 2
  • Satoshi Maruyama
    • 1
    • 3
  • Tatsuya Abé
    • 1
    • 3
  • Takanori Kobayashi
    • 1
  • Manabu Yamazaki
    • 1
  • Akinori Funayama
    • 1
    • 2
  • Susumu Shingaki
    • 2
  • Tadaharu Kobayashi
    • 2
  • Cheng Jun
    • 1
  • Takashi Saku
    • 1
    • 3
    Email author
  1. 1.Division of Oral Pathology, Department of Tissue Regeneration and ReconstructionNiigata University Graduate School of Medical and Dental SciencesNiigataJapan
  2. 2.Division of Reconstructive Surgery for Oral and Maxillofacial Region, Department of Tissue Regeneration and ReconstructionNiigata University Graduate School of Medical and Dental SciencesNiigataJapan
  3. 3.Oral Pathology Section, Department of Surgical PathologyNiigata University HospitalNiigataJapan

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