Clin Exp Otorhinolaryngol.  2017 Sep;10(3):213-220. 10.21053/ceo.2016.01263.

Cytokeratin 13, Cytokeratin 17, and Ki-67 Expression in Human Acquired Cholesteatoma and Their Correlation With Its Destructive Capacity

• Hamed MA
• Nakata S
• Shiogama K
• Suzuki K
• Sayed RH
• Nishimura
• Iwata N
• Sakurai K
• Badawy BS
• Inada K
• Tsuge H
• Tsutsumi Y

• KMID: 2395734
• DOI: http://doi.org/10.21053/ceo.2016.01263

Abstract

OBJECTIVES
Cholesteatoma is a nonneoplastic destructive lesion of the temporal bone with debated pathogenesis and bone resorptive mechanism. Both molecular and cellular events chiefly master its activity. Continued research is necessary to clarify factors related to its aggressiveness. We aimed to investigate the expression of Ki-67, cytokeratin 13 (CK13) and cytokeratin 17 (CK17) in acquired nonrecurrent human cholesteatoma and correlate them with its bone destructive capacity.
METHODS
A prospective quantitative immunohistochemical study was carried out using fresh acquired cholesteatoma tissues (n=19), collected during cholesteatoma surgery. Deep meatal skin tissues from the same patients were used as control (n=8). Cholesteatoma patients were divided into 2 groups and compared (invasive and noninvasive) according to a grading score for bone resorption based upon clinical, radiologic and intraoperative findings. To our knowledge, the role of CK17 in cholesteatoma aggressiveness was first investigated in this paper.
RESULTS
Both Ki-67 and CK17 were significantly overexpressed in cholesteatoma than control tissues (P < 0.001 for both Ki-67 and CK17). In addition, Ki-67 and CK17 were significantly higher in the invasive group than noninvasive group of cholesteatoma (P=0.029, P=0.033, respectively). Furthermore, Ki-67 and CK17 showed a moderate positive correlation with bone erosion scores (r=0.547, P=0.015 and r=0.588, P=0.008, respectively). In terms of CK13, no significant difference was found between cholesteatoma and skin (P=0.766).
CONCLUSION
Both Ki-67 and CK17 were overexpressed in cholesteatoma tissue and positively correlated with bone resorption activity. The concept that Ki-67 can be a predictor for aggressiveness of cholesteatoma was supported. In addition, this is the first study demonstrating CK17 as a favoring marker in the aggressiveness of acquired cholesteatoma.

Keyword

Cholesteatoma; Middle Ear; Bone Resorption; Ki-67 Antigen; Cytokeratin 17

MeSH Terms

Bone Resorption
Cholesteatoma*
Ear, Middle
Humans*
Keratin-13*
Keratin-17*
Keratins*
Ki-67 Antigen
Prospective Studies
Skin
Temporal Bone
Keratin-13
Keratin-17
Keratins
Ki-67 Antigen

Figure

• 

  Fig. 1. (A, B) Expression of Ki-67 and cytokeratin 17 (CK17) in cholesteatoma versus skin (tissue group, mean±SE) and (C, D) in invasive versus non-invasive cholesteatoma (cholesteatoma group, mean±SE). Note overexpression of both Ki-67 and CK17 in cholesteatoma compared to skin tissues (P<0.001), and in invasive group than noninvasive one (P<0.05). Independent t-test. SE, standard error of the mean.

• 

  Fig. 2. Spearman correlation of (A) Ki-67 and (B) cytokeratin 17 (CK 17) with scores of bone erosion. Moderate positive correlations were observed between Ki-67 (r=0.547, P<0.05) and CK17 (r=0.588, P<0.01) and grading score used in our study.

• 

  Fig. 3. Expression of (A) Ki-67 in invasive, (B) Ki-67 in noninvasive, (C) cytokeratin 17 (CK17) in invasive, and (D) CK17 in noninvasive cholesteatoma. Overexpression of Ki-67 is observed in the invasive group as compared to the noninvasive one (arrows). The same is true for CK17 (arrows) (Mayer’s hematoxylin, ×200).

• 

  Fig. 4. Expression of (A) cytokeratin 13 (CK13) and (B) CK17 in skin, and (C) CK13 and (D) CK17 in cholesteatoma tissues. Expression of CK17 was much increased in cholesteatoma (arrows) compared to the skin but CK13 did not show much difference in both skin and cholesteatoma tissues (Mayer’s hematoxylin, ×200).

Reference

1. Olszewska E, Wagner M, Bernal-Sprekelsen M, Ebmeyer J, Dazert S, Hildmann H, et al. Etiopathogenesis of cholesteatoma. Eur Arch Otorhinolaryngol. 2004; Jan. 261(1):6–24.

2. Bujia J, Schilling V, Holly A, Stammberger M, Kastenbauer E. Hyperproliferation-associated keratin expression in human middle ear cholesteatoma. Acta Otolaryngol. 1993; May. 113(3):364–8.

3. Juhn SK, Jung MK, Hoffman MD, Drew BR, Preciado DA, Sausen NJ, et al. The role of inflammatory mediators in the pathogenesis of otitis media and sequelae. Clin Exp Otorhinolaryngol. 2008; Sep. 1(3):117–38.

4. Peek FA, Huisman MA, Berckmans RJ, Sturk A, Van Loon J, Grote JJ. Lipopolysaccharide concentration and bone resorption in cholesteatoma. Otol Neurotol. 2003; Sep. 24(5):709–13.

5. Olszewska E, Jakimowicz-Rudy J, Knas M, Chilimoniuk M, Pietruski JK, Sieskiewicz A. Cholesteatoma-associated pathogenicity: potential role of lysosomal exoglycosidases. Otol Neurotol. 2012; Jun. 33(4):596–603.

6. Sikka K, Sharma SC, Thakar A, Dattagupta S. Evaluation of epithelial proliferation in paediatric and adult cholesteatomas using the Ki-67 proliferation marker. J Laryngol Otol. 2012; May. 126(5):460–3.

7. Mallet Y, Nouwen J, Lecomte-Houcke M, Desaulty A. Aggressiveness and quantification of epithelial proliferation of middle ear cholesteatoma by MIB1. Laryngoscope. 2003; Feb. 113(2):328–31.

8. Banco B, Grieco V, Di Giancamillo M, Greci V, Travetti O, Martino P, et al. Canine aural cholesteatoma: a histological and immunohistochemical study. Vet J. 2014; Jun. 200(3):440–5.

9. Shiwa M, Kojima H, Moriyama H. Expression of transforming growth factor-alpha (TGF-alpha) in cholesteatoma. J Laryngol Otol. 1998; Aug. 112(8):750–4.

10. Yamamoto-Fukuda T, Takahashi H, Terakado M, Hishikawa Y, Koji T. Expression of keratinocyte growth factor and its receptor in noncholesteatomatous and cholesteatomatous chronic otitis media. Otol Neurotol. 2010; Jul. 31(5):745–51.

11. Macias MP, Gerkin RD, Macias JD. Increased amphiregulin expression as a biomarker of cholesteatoma activity. Laryngoscope. 2010; Nov. 120(11):2258–63.

12. Lee SH, Jang YH, Tae K, Park YW, Kang MJ, Kim KR, et al. Telomerase activity and cell proliferation index in cholesteatoma. Acta Otolaryngol. 2005; Jul. 125(7):707–12.

13. Yetiser S, Satar B, Aydin N. Expression of epidermal growth factor, tumor necrosis factor-alpha, and interleukin-1alpha in chronic otitis media with or without cholesteatoma. Otol Neurotol. 2002; Sep. 23(5):647–52.

14. Juhasz A, Sziklai I, Rakosy Z, Ecsedi S, Adany R, Balazs M. Elevated level of tenascin and matrix metalloproteinase 9 correlates with the bone destruction capacity of cholesteatomas. Otol Neurotol. 2009; Jun. 30(4):559–65.

15. Oger M, Alpay HC, Orhan I, Onalan EE, Yanilmaz M, Sapmaz E. The effect of BMP-2, BMP-4 and BMP-6 on bone destruction of cholesteatoma presence. Am J Otolaryngol. 2013; Nov-Dec. 34(6):652–7.

16. Kuo CL. Etiopathogenesis of acquired cholesteatoma: prominent theories and recent advances in biomolecular research. Laryngoscope. 2015; Jan. 125(1):234–40.

17. Vennix PP, Kuijpers W, Peters TA, Tonnaer EL, Ramaekers FC. Epidermal differentiation in the human external auditory meatus. Laryngoscope. 1996; Apr. 106(4):470–5.

18. Vennix PP, Kuijpers W, Peters TA, Tonnaer EL, Ramaekers FC. Keratinocyte differentiation in acquired cholesteatoma and perforated tympanic membranes. Arch Otolaryngol Head Neck Surg. 1996; Aug. 122(8):825–32.

19. Sudhoff H, Bujia J, Fisseler-Eckhoff A, Holly A, Schulz-Flake C, Hildmann H. Expression of a cell-cycle-associated nuclear antigen (MIB 1) in cholesteatoma and auditory meatal skin. Laryngoscope. 1995; Nov. 105(11):1227–31.

20. Raynov AM, Moon SK, Choung YH, Hong SP, Park K. Nucleoplasm staining patterns and cell cycle-associated expression of Ki-67 in middle ear cholesteatoma. Am J Otolaryngol. 2005; Sep-Oct. 26(5):296–301.

21. Akdogan V, Yilmaz I, Canpolat T, Ozluoglu LN. Role of Langerhans cells, Ki-67 protein and apoptosis in acquired cholesteatoma: prospective clinical study. J Laryngol Otol. 2013; Mar. 127(3):252–9.

22. Chung JH, Lee SH, Park CW, Kim KR, Tae K, Kang SH, et al. Expression of apoptotic vs antiapoptotic proteins in middle ear cholesteatoma. Otolaryngol Head Neck Surg. 2015; Dec. 153(6):1024–30.

23. Kuczkowski J, Pawelczyk T, Bakowska A, Narozny W, Mikaszewski B. Expression patterns of Ki-67 and telomerase activity in middle ear cholesteatoma. Otol Neurotol. 2007; Feb. 28(2):204–7.

24. Aslier M, Erdag TK, Sarioglu S, Guneri EA, Ikiz AO, Uzun E, et al. Analysis of histopathological aspects and bone destruction characteristics in acquired middle ear cholesteatoma of pediatric and adult patients. Int J Pediatr Otorhinolaryngol. 2016; Mar. 82:73–7.

25. Kim HJ, Tinling SP, Chole RA. Increased proliferation and migration of epithelium in advancing experimental cholesteatomas. Otol Neurotol. 2002; Nov. 23(6):840–4.

26. Lee RJ, Sidey C, Narula AA, James RF. The nature of the epithelium in acquired cholesteatoma: part 3: cytokeratin patterns in aural epithelial and cholesteatoma cells grown in cell culture. Clin Otolaryngol Allied Sci. 1994; Dec. 19(6):516–20.

27. Toyoshima T, Vairaktaris E, Nkenke E, Schlegel KA, Neukam FW, Ries J. Cytokeratin 17 mRNA expression has potential for diagnostic marker of oral squamous cell carcinoma. J Cancer Res Clin Oncol. 2008; Apr. 134(4):515–21.

28. Mikami T, Cheng J, Maruyama S, Kobayashi T, Funayama A, Yamazaki M, et al. Emergence of keratin 17 vs. loss of keratin 13: their reciprocal immunohistochemical profiles in oral carcinoma in situ. Oral Oncol. 2011; Jun. 47(6):497–503.

29. Jin L, Wang G. Keratin 17: a critical player in the pathogenesis of psoriasis. Med Res Rev. 2014; Mar. 34(2):438–54.

30. Desloge RB, Carew JF, Finstad CL, Steiner MG, Sassoon J, Levenson MJ, et al. DNA analysis of human cholesteatomas. Am J Otol. 1997; Mar. 18(2):155–9.