Hakan BİRKENT, Burcu ESER, Hamdi TAŞLI, Burak AŞIK, Mustafa GEREK

Larenks Skuamöz Hücreli Karsinom Örneklerinde Pepsin Araştırılması

Amaç: Skuamöz hücreli karsinom (SHK) larenks kanserlerinin % 95'ini oluşturur. Son yirmi yılda SHK riskinin artmasında kabul gören faktörlerden biri larengofaringeal reflü hastalığıdır (LFR). Larenks kanserinde LFR'nin rolü ve mekanizması belirsizliğini korumaktadır. Bu çalışmanın amacı, larenks kanseri gelişiminde LFR'nin rol oynadığı hipotezini test etmek için larengeal formalinle fikse edilmiş parafine gömülü (FFPG) skuamöz hücreli karsinom (SHK) örneklerinde pepsin varlığını araştırmaktır. Yöntemler: Total larenjektomi uygulanan ileri evre larengeal kanser hastalarının toplam 29 patoloji örneği değerlendirildi. Patoloji bölümünde parafin bloklarında tutulan örneklerin histopatolojik olarak SHK olduğu doğrulandı. Pepsin enzim seviyelerini ölçmek için western blot (WB) veya enzime bağlı immunosorbent assay (ELISA) testleri kullanıldı. Bulgular: WB ve ELISA test sonuçlarına gore, 29 formalinle fikse edilmiş, parafine gömülü (FFPG) larengeal SHK örneklerinin hiçbirinde pepsin enzimine rastlanmadı. Bununla birlikte, pozitif kontrollerde hem WB hem de ELISA testleri ile pepsin enzimi tespit edildi. Sonuçlar: Bu çalışmada malign larengeal skuamöz hücreli karsinom örneklerinde pepsine rastlanmadı. LFR ve larengeal SHK arasındaki ilişki hala belirsizliğini korumakla birlikte larengeal SHK'da pepsin ve LFR'nin rolünü doğrulamak için daha ileri çalışmalara ihtiyaç duyulmaktadır.

Investigation of Pepsin in Laryngeal Squamous Cell Carcinoma Specimens

Objective: Squamous cell carcinoma (SCC) composes 95% of the carcinoma of the larynx. One of the factor recognizedin the past two decades as increasing the risk of SCC is laryngopharyngeal reflux (LPR). The role and mechanism ofLPR in laryngeal cancer remain unclear. The aim of this study was to assess the presence of pepsin in laryngealformalin-fixed, paraffin-embedded (FFPE) squamous cell carcinoma (SCC) specimens in order to test the hypothesisthat LPR plays a role in the development of laryngeal SCC.Method: A total of 29 pathology specimens of advanced-stage laryngeal cancer patients who had undergone totallaryngectomy were evaluated. The specimens were confirmed histopathologically as SCC and maintained in paraffinblocks in the pathology department. The western blot (WB) or the enzyme-linked immunosorbent assay (ELISA)analyses were used to measure pepsin enzyme levels.Results: According to the WB and ELISA analyses, there was no presence of the pepsin enzyme in any of the 29formalin-fixed, paraffin-embedded (FFPE) laryngeal SCC specimens. However, pepsin was detected with both the WBand the ELISA tests in positive controls and fresh rat stomach specimen.Conclusion: Pepsin was not present in any of the malignant laryngeal squamous cell carcinoma specimens in thisstudy. Relationship between LPR and laryngeal SCC still remains unclear and further studies are needed to verify therole of pepsin and LPR in laryngeal SCC.

PDF

___

1.Cann CI, Fried MP, Rothman KJ. Epidemiology of squamous cell cancer of the head and neck. Otolaryngol Clin North Am. 1985; 18: 367-88.
2. Landis SH, Murray T, Bolden S, et al. Cancer statistics, 1998. CA Cancer J Clin. 1998; 48: 6-29.
3. Berrino F. Survival of cancer patients in Europe: the EUROCARE-2 study. IARC Sci Publ. 1999; 151: 1-572.
4. Rothman KJ, Cann CI, Flanders D, et al. Epidemiology of laryngeal cancer. Epidemiologic reviews. 1980; 2: 195– 209.
5. Copper MP, Smit CF, Stanojcic LD, et al. High incidence of laryngopharyngeal reflux in patients with head and neck cancer. Laryngoscope. 2000; 110: 1007–11.
6. Samuels TL, Johnston N. Pepsin as a Marker of Extraesophageal Reflux. Ann Otol Rhinol Laryngol. 2010; 119: 203-8.
7. Wassenaar E, Johnston N, Merati A, et al. Pepsin detection in patients with laryngopharyngeal reflux before and after fundoplication. Surg Endosc. 2011; 25: 3870-6.
8. Glanz H, Kleinsasser O. Chronic laryngitis and carcinoma. Arch Otorhinolaryngol. 1976; 212: 57-75.
9. Johnston N, Yan JC, Hoekzema CR, et al. Pepsin promotes proliferation of laryngeal and pharyngeal epithelial cells. Laryngoscope. 2012; 122: 1317-25.
10. Qadeer MA, Colabianchi N, Vaezi MF. Is GERD a risk factor for laryngeal cancer?. Laryngoscope. 2005; 115: 486–91.
11. Belafsky PC, Postma GN, Koufman JA. The validity and reliability of the reflux finding score (RFS). Laryngoscope. 2001; 111: 1313-7.
12. Tasli H, Eser B, Asik MB, et al. Does Pepsin Play a Role in Etiology of Laryngeal Nodules?. J Voice. 2018 May 11. pii: S0892-1997(18)30041-9.
13. Johnston N, Dettmar PW, Bishwokarma B, et al. Activity/stability of human pepsin: implications for reflux attributed laryngeal disease. Laryngoscope. 2007; 117: 1036–9.
14. Johnston N, Wells CW, Samuels TL, et al. Rationale for targeting pepsin in the treatment of reflux disease. Ann Otol Rhinol Laryngol. 2010; 119: 547–58.
15. Johnston N, Peter W. Dettmar PW, et al. Laryngopharyngeal reflux and GERD. Ann N Y Acad Sci. 2013; 1300: 71-9.
16. Coca-Pelaz A, Rodrigo JP, Takes RP, et al. Relationship between reflux and laryngeal cancer. Head Neck. 2013; 35: 1814-8.
17. Nilsson M, Chow WH, Lindblad M, et al. No association between gastroesophageal reflux and cancers of the larynx and pharynx. Cancer Epidemiol Biomarkers Prev. 2005; 14: 1194-7.
18. Francis DO, Maynard C, Weymuller EA, et al. Reevaluation of gastroesophageal reflux disease as a risk factor for laryngeal cancer. Laryngoscope. 2011; 121: 102-5.
19. Ozlugedik S, Yorulmaz I, Gokcan K. Is laryngopharyngeal reflux an important risk factor in the development of laryngeal carcinoma?. Eur Arch Otorhinolaryngol. 2006; 263: 339–43.
20. Geterud A, Bove M, Ruth M. Hypopharyngeal acid exposure: an independent risk factor for laryngeal cancer?. Laryngoscope. 2003; 113: 2201–5.
21. el-Serag HB, Sonnenberg A. Comorbid occurrence of laryngeal or pulmonary disease with esophagitis in United Steates militery veterans. Gastroenterology. 1997; 113: 755-60.
22. Qadeer MA, Colabianchi N, Vaezi MF. Is GERD a risk factor for laryngeal cancer?. Laryngoscope. 2005; 115: 486–91.
23. Galli J, Cammarota G, Calò L, et al. The role of acid and alkaline reflux in laryngeal squamous cell carcinoma. Laryngoscope. 2002; 112: 1861–5.
24. Kim TH, Lee KJ, Yeo M, et al. Pepsin detection in the sputum/saliva for the diagnosis of gastroesophageal reflux disease in patients with clinically suspected atypical gastroesophageal reflux disease symptoms. Digestion. 2008; 77: 201-6.
25. Johnston N, Yan JC, Hoekzema CR, et al. Pepsin promotes proliferation in normal and transformed laryngopharyngeal epithelial cells. Laryngoscope. 2012; 122: 1317–25.
26. Shi SR, Key ME, Kalra KL. Antigen retrieval in formalin fixed, paraffin-embedded tissues: an enhancement method for immunohistochemical staining based on microwave oven heating of tissue sections. J Histochem Cytochem. 1991; 39: 741–8
27. Tanca A, Pagnozzi D, Addis MF. Setting proteins free: progresses and achievements in proteomics of formalin-fixed, paraffin-embedded tissues. Proteomics Clin Appl. 2011; 6: 7–21.
28. Ostasiewicz P, Zielinska DF, Mann M, et al. Proteome, phosphoproteome, and N-glycoproteome are quantitatively preserved in formalin-fixed paraffinembedded tissue and analyzable by high-resolution mass spectrometry. J Proteome Res. 2010; 9: 3688– 700.
29. Balgley BM, Guo T, Zhao K, et al. Evaluation of archival time on shotgun proteomics of formalin-fixed and paraffin-embedded tissues. J Proteome Res. 2009; 8: 917-25.
30. Vaezi ME, Schroeder PL, Richter JE. Reproducibility of proximal probe pH parameters in 24-hour ambulatory esophageal pH monitoring. Am J Gastroenterol. 1997; 92: 825-9.