Head and Neck Pathology

, Volume 4, Issue 4, pp 282–289 | Cite as

Analysis of Immunohistochemical Expression of K19 in Oral Epithelial Dysplasia and Oral Squamous Cell Carcinoma Using Color Deconvolution-Image Analysis Method

  • Rima A. SafadiEmail author
  • Atika S. Musleh
  • Taiseer H. Al-Khateeb
  • Abed Al-Hadi Hamasha
Original Paper


K19 is an intermediate filament protein that has been investigated in oral squamous cell carcinoma (OSCC), but that has not been correlated with the amount of keratin produced within well-differentiated OSCC grade. The aim of the present study was to objectively analyze K19 immunoexpression in OSCC and to validate the utility of K19 in differentiation among grades of oral epithelial dysplasia (OED). Formalin-fixed tissues of 36 primary OSCC (22 well, 10 moderately, 4 poorly differentiated), 43 OED (23 mild, 8 moderate, 12 severe), and 11 normal oral epithelium (NOE) were included. K19 was immunostained using HRP-DAB method. The percentage of K19-positive area was found using color deconvolution program in ImageJ® image analysis system (public domain software, National Institutes of Health, Bethesda, MD, USA) and analyzed using independent samples t tests and ANOVA test. K19 scores in NOE, mild, moderate and severe OED were: 1.8, 3.4, 21, and 50.3%, respectively, with significant association with the grade (t test P < 0.05). Well-differentiated OSCC with <30% keratin pearl formation expressed significantly higher K19 scores compared to well-differentiated OSCC with >30% keratin pearls (28.6 and 1.2%, respectively, P < 0.05). K19 scores in moderately and poorly differentiated OSCC were 60.8 and 61.3%, respectively. K19 scores significantly differentiated between two subgroups of tumors within well-differentiated OSCC grade and reflected histologic differentiation as well as probably predicting the clinical outcome. Combining K19 immunostain with the regular H&E stain may be helpful to facilitate and assure assigning a more accurate grade for OED.


Oral squamous cell carcinoma Epithelial dysplasia Immunohistochemistry K19 Image analysis 


  1. 1.
    De Vicente J, Recio O, Pendás S, et al. Oral squamous cell carcinoma of the mandibular region A survival study. Head Neck. 2001;23(7):536–43.CrossRefPubMedGoogle Scholar
  2. 2.
    Warnakulasuriya S. Global epidemiology of oral and oropharyngeal cancer. Oral Oncol. 2009;45(4–5):309–16.CrossRefPubMedGoogle Scholar
  3. 3.
    Silverman SJ, Sugerman P. Oral premalignancies and squamous cell carcinoma. Clin Dermatol. 2000;18(5):563–8.CrossRefPubMedGoogle Scholar
  4. 4.
    Kramer I, Lucas R, Pindborg J, et al. Definition of leukoplakia and related lesions: An aid to studies on oral precancer. Oral Surg Oral Med Oral Pathol. 1978;46(4):518–39.CrossRefPubMedGoogle Scholar
  5. 5.
    Silverman SJ, Gorsky M, Kaugars G. Leukoplakia, dysplasia, and malignant transformation. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 1996;82(2):117.CrossRefPubMedGoogle Scholar
  6. 6.
    Abbey L, Kaugars G, Gunsolley J, et al. Intraexaminer and interexaminer reliability in the diagnosis of oral epithelial dysplasia. Oral Pathol Oral Radiol Endod. 1995;80(2):188–91.CrossRefGoogle Scholar
  7. 7.
    Brothwell DJ, Lewis DW, Bradley G, et al. Observer agreement in the grading of oral epithelial dysplasia. Community Dent Oral Epidemiol. 2003;31(4):300–5.CrossRefPubMedGoogle Scholar
  8. 8.
    Fischer DJ, Epstein JB, Morton TH, et al. Interobserver reliability in the histopathologic diagnosis of oral pre-malignant and malignant lesions. J Oral Pathol Med. 2004;33(2):65–70.CrossRefPubMedGoogle Scholar
  9. 9.
    Hu L, Crowe DL, Rheinwald JG, et al. Abnormal expression of retinoic acid receptors and keratin 19 by human oral and epidermal squamous cell carcinoma cell lines. Cancer Res. 1991;51(15):3972–81.PubMedGoogle Scholar
  10. 10.
    Takeda T, Sugihara K, Hirayama Y, et al. Immunohistological evaluation of Ki-67, p63, CK19 and p53 expression in oral epithelial dysplasias. J Oral Pathol Med. 2006;35(6):369–75.CrossRefPubMedGoogle Scholar
  11. 11.
    Warnakulasuriya S. Lack of molecular markers to predict malignant potential of oral precancer. J Pathol. 2000;190(4):407–9.CrossRefPubMedGoogle Scholar
  12. 12.
    Fradette J, Germain L, Seshaiah P, et al. The type I keratin 19 possesses distinct and context-dependent assembly properties. J Biol Chem. 1998;273(52):35176–84.CrossRefPubMedGoogle Scholar
  13. 13.
    Michel M, Torok N, Godbout MJ, et al. Keratin 19 as a biochemical marker of skin stem cells in vivo and in vitro: Keratin 19 expressing cells are differentially localized in function of anatomic sites, and their number varies with donor age and culture stage. J Cell Sci. 1996;109(Pt5):1017–70.PubMedGoogle Scholar
  14. 14.
    Lindberg K, Rheinwald JG. Suprabasal 40kd keratin (K19) expression as an immunohistologic marker of premalignancy in oral epithelium. Am J Pathol. 1989;134(1):89–98.PubMedGoogle Scholar
  15. 15.
    Zhong LP, Chen WT, Zhang CP, et al. Increased CK19 expression correlated with pathologic differentiation grade and prognosis in oral squamous cell carcinoma patients. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2007;104(3):377–84.CrossRefPubMedGoogle Scholar
  16. 16.
    Anneroth G, Hansen LS. A methodologic study of histologic classification and grading of malignancy in oral squamous cell carcinoma. Scand J Dent Res. 1984;92(5):448–68.PubMedGoogle Scholar
  17. 17.
    Ruifrok AC, Johnston DA. Quantification of histochemical staining by color deconvolution. Anal Quant Cytol Histol. 2001;23(4):291–9.PubMedGoogle Scholar
  18. 18.
    Elias JM, Gown AM, Nakamura RM, et al. Quality control in immunohistochemistry. Report of a workshop sponsored by the Biological Stain Commission. Am J Clin Pathol. 1989;92(6):836–43.PubMedGoogle Scholar
  19. 19.
    Fillies T, Jogschies M, Kleinheinz J, et al. Cytokeratin alteration in oral leukoplakia and oral squamous cell carcinoma. Oncol Rep. 2007;18(3):639–43.PubMedGoogle Scholar
  20. 20.
    Nie M, Zhong L, Zeng G, et al. The changes of cytokeratin 19 during oral carcinogenesis. Zhonghua Kou Qiang Yi Xue Za Zhi. 2002;37(3):187–90.PubMedGoogle Scholar
  21. 21.
    Küffer R, Lombardi T. Premalignant lesions of the oral mucosa. A discussion about the place of oral intraepithelial neoplasia (OIN). Oral Oncol. 2002;38(2):125–30.CrossRefPubMedGoogle Scholar
  22. 22.
    Warnakulasuriya S, Reibel J, Bouquot J, et al. Oral epithelial dysplasia classification systems: Predictive value, utility, weaknesses and scope for improvement. J Oral Pathol Med. 2008;37(3):127–33.CrossRefPubMedGoogle Scholar
  23. 23.
    Kujan O, Oliver RJ, Khattab A, et al. Evaluation of a new binary system of grading oral epithelial dysplasia for prediction of malignant transformation. Oral Oncol. 2006;42(10):987–93.CrossRefPubMedGoogle Scholar
  24. 24.
    Su L, Morgan PR, Thomas JA, et al. Expression of keratin 14 and 19 mRNA and protein in normal oral epithelia, hairy leukoplakia, tongue biting and white sponge nevus. J Oral Pathol Med. 1993;22(4):183–9.CrossRefPubMedGoogle Scholar
  25. 25.
    Toyoshima T, Vairaktaris E, Nkenke E, et al. Cytokeratin 17 mRNA expression has potential for diagnostic marker of oral squamous cell carcinoma. J Cancer Res Clin Oncol Erratum in: J Cancer Res Clin Oncol. 2008;34(4):523-24,134(4):515-21.Google Scholar
  26. 26.
    Fillies T, Werkmeister R, Packeisen J, et al. Cytokeratin 8/18 expression indicates a poor prognosis in squamous cell carcinomas of the oral cavity. BMC Cancer. 2006;13(6):10.CrossRefGoogle Scholar
  27. 27.
    Yoshida Y. [Effective biological marker to detect oral squamous cancer cells–expression patterns of CK 10, 17, 19 and SCCA mRNA–]. Kokubyo, Gakkai Zasshi. 2007;73 74 (3,1):37-42.Google Scholar
  28. 28.
    Bryne M. Is the invasive front of an oral carcinoma the most important area for prognostication? Oral Dis. 1998;4(2):70–7.CrossRefPubMedGoogle Scholar

Copyright information

© Humana 2010

Authors and Affiliations

  • Rima A. Safadi
    • 1
    • 2
    Email author
  • Atika S. Musleh
    • 3
  • Taiseer H. Al-Khateeb
    • 1
    • 2
  • Abed Al-Hadi Hamasha
    • 4
  1. 1.Department of Oral Medicine and Surgery, Faculty of DentistryJordan University of Science and TechnologyIrbidJordan
  2. 2.King Abdullah University HospitalIrbidJordan
  3. 3.Ministry of HealthAmmanJordan
  4. 4.Department of Preventive Dentistry, Faculty of DentistryJordan University of Science and TechnologyIrbidJordan

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