Sığır melanomlarında S100, Vimentin, PCNA, p53 ve MMP-9 ekspresyonlarının immunohistokimyasal olarak değerlendirilmesi

Year 2021, Volume: 10 Issue: 1, 43 - 49, 22.06.2021

Emin KARAKURT Uğur AYDIN Enver BEYTUT Engin KILIÇ Serpil DAĞ Hilmi NUHOĞLU Uğur YILDIZ Ayfer YILDIZ Emre KURTBAŞ

https://doi.org/10.31196/huvfd.859433

Abstract

Bu çalışmada, sığır melanomlarının agresifliğini ve tanısı için PCNA, p53, MMP-9, Vimentin ve S100 ekspresyonlarının immünohistokimyasal olarak değerlendirilmesi amaçlanmıştır. Bu çalışmanın materyali bölümümüze getirilen 10 sığırdan alınan melanom biyopsi örneklerinden oluşmuştur. Sığırlardan alınan doku örnekleri, %10’luk tamponlu formalin solüsyonunda tespit edildi. Rutin prosedürlerden sonra hazırlanan parafin bloklardan 5 μm kalınlığında kesitler alındı. Ağartma için bazı ağır pigmentli tümör kesitleri deparafinize edildi, hidratlandı ve % 10’luk hidrojen peroksit (H2O2) çözeltisi içinde 65oC'de 5 saat süreyle, kesitler berrak hale gelene kadar inkübe edildi ve kesitlere histopatolojik değişikliklerin tespit edilmesi için Hematoksilen & Eozin (H&E) boyaması yapıldı. Kesitler ışık mikroskobu altında incelendi ve fotoğraflandı. İmmünohistokimyasal yöntem olarak Avidin-Biotin Peroksidaz yöntemi kullanıldı. Tümöral kitlenin soliter, sert, tüysüz, oval-yuvarlak şekilli ve oldukça büyük olduğunu gözlemledik. Sitoplazmalarında çok sayıda büyük kahverengimsi siyah granüler melanin pigmenti içeren iğsi ve epiteloid tipi tümör hücrelerini tespit ettik. Tüm melanom vakaları S100, Vimentin, PCNA, p53 ve MMP-9 ekspresyonları için immün pozitifti. Sonuç olarak, bu immünohistokimyasal belirteçlerin sığır melanomlarının prognozunda ve teşhisinde oldukça kullanışlı olduğunu düşünmekteyiz.

Keywords

immunohistokimya, melanom, prognoz, sığır

Immunohistochemical assessment of S100, Vimentin, PCNA, p53 and MMP-9 expressions in bovine melanomas

Abstract

This study aimed to evaluate the expression of PCNA, p53, MMP-9, Vimentin and S100 immunohistochemically and determine the aggressiveness in diagnosis of bovine melanomas. The material of this study consisted of melanoma biopsy samples taken from 10 cattle brought to our department. Tissue samples from cattle were fixed in 10% buffered formalin solution. After routine procedures paraffin blocks were cut sections of 5 μm thickness. For bleaching, some heavily pigmented tumour sections were deparaffinized, hydrated, and incubated in 10% solution of hydrogen peroxide (H2O2) for 5 hours at 65oC until sections appeared clear and Hematoxylin & Eosin (H&E) staining was applied to the sections to detect histopathological changes. Sections were examined and photographed under a light microscope. Avidin-Biotin Peroxidase was used as the immunohistochemical method. We observed that the tumoral mass was solitary, firm, hairless, oval-round shaped and quite large. We detected spindle and epithelioid type tumoral cells containing a lot of large brownish-black granular melanin pigments in their cytoplasm. All melanoma cases were immune positive for S100, Vimentin, PCNA, p53 and MMP-9 expressions. In conclusion, we think that these immunohistochemical markers are quite convenient in evaluating the prognosis and diagnosis of bovine melanomas.

Keywords

bovine, immunohistochemistry, melanoma, prognosis

References

• Referans1Aksenenko MB, Ruksha TG, 2013: Analysis of the application of MMP-9 inhibitor in skin melanoma: experimental study. Bull Exp Biol Med, 154 (5), 594-596.
• Referans2Babić T, Grabarević Ž, Vuković S, Kos J, Matičić D, 2009: Congenital melanoma in a 3-month old bull calf - a case report. Veterinarski Arhiv, 79 (4), 315-320.
• Referans3Beytut E, Kılıç E, Yayla S, 2018: Histopathological and immunohistochemical evaluation of congenital cutaneous melanomas in calves (3 cases). Ankara Üniv Vet Fak Derg, 65 (4), 425-432.
• Referans4Bhadaniya AR, Trangadia BJ, Prasad MC, Fefar DT, Kalaria VA, Vadaliya JV, Modi KS, 2015: Pathology of melanocytic tumour in cattle. Indian J Vet Pathol, 39 (3), 249-250.
• Referans5Brito MF, França TN, Jabour F, Seixas JN, Andrade G, Oliveira LI, Peixoto, P, 2009: Metastasizing oral melanoma in a cow. Ciencia Rural, 39 (4), 1236-1240.
• Referans6Chandrashekaraiah GB, Ballari SV, Manjunatha K, Chavadhal N, Radder SK, 2014: Malignant melanoma in a Hallikar bullock. Int J Vet Sci, 3 (2), 65-67.
• Referans7Garma- Aviña A, Valli VE, Lumsden JH, 1981: Cutaneous melanomas in domestic animals. J Cutan Pathol, 8 (1), 3-24.
• Referans8Hemanth I, Amaravathi P, Anand Kumar A, Devaratnam J, Bharathi S, Sailaja N, Kamalakar G, Sasidhar Babu N, 2014: Cutaneous melano-fibroma in a bullock – a rare concurrence of melanoma and fibroma. Int J Sci Environ Technol, 3 (2), 659-662.
• Referans9Javanbakht J, Sasani F, Adibhashemi F, Hemmati S, 2014: Comparative histopathological diagnosis of cutaneous melanoma by H&E, special staining and immunohistochemical methods against cutaneous squamous cell carcinoma in horse and bovine. J Bioanal and Biomed, 6 (4), 19-23.
• Referans10Koenig A, Wojcieszyn J, Weeks BR, Modiano JF, 2001: Expression of S100a, vimentin, NSE, and melan A/MART-1 in seven canine melanoma cells lines and twenty-nine retrospective cases of canine melanoma. Vet Pathol, 38 (4), 427-435.
• Referans11Madheswaran R, Shahana S, Gopal K, Sankar P, 2019: A case report of non-systemic highly aggressive melanoma in a cow. Indian J Vet Pathol, 43 (2), 124-126.
• Referans12Mesarić M, Zadnik T, Cerne M, 2002: Malignant melanoma in a cow. Acta Vet-Beograd, 52 (1), 59-64.
• Referans13Miller MA, Weaver AD, Stogsdill PL, Fischer JR, Kreeger JM, Nelson SL, Turk JR, 1995: Cutaneous melanocytomas in 10 young cattle. Vet Pathol, 32 (5), 479-484.
• Referans14Naveen B, Chandrashekar M, Vishwaradhya TM, Shambulinga M, Kumar P, Singh D, Rao S, 2013: Malignant melanoma in indigenous cattle - a case report. Haryana Vet, 52, 135-136.
• Referans15Ozyildiz Z, Ceylan C, Yilmaz R, Ozsoy SY, 2012: Immunohistochemical characterization of perineal melanoma in Kilis goats. Biotech Histochem, 87 (6), 408-412.
• Referans16Pazhanivel N, Ezakial Napolean, Murali Manohar B, Ravi U, 2003: A case of cutaneous melanoma in a bull. Indian J Anim Res, 37 (2), 151-152.
• Referans17Qin J, Li S, Zhang C, Gao DW, Li Q, Zhang H, Jin XD, Liu Y, 2017: Apoptosis and injuries of heavy ion beam and x-ray radiation on malignant melanoma cell. Exp Biol Med (Maywood), 242 (9), 953-960.
• Referans18Ramos-Vara JA, Frank CB, DuSold D, Miller MA, 2014: Immunohistochemical expression of melanocytic antigen PNL2, Melan A, S100, and PGP 9.5 in equine melanocytic neoplasms. Vet Pathol, 51 (1), 161-166.
• Referans19Ramos-Vara JA, Miller MA, 2011: Immunohistochemical identification of canine melanocytic neoplasms with antibodies to melanocytic antigen PNL2 and tyrosinase: comparison with Melan A. Vet Pathol, 48 (2), 443-450.
• Referans20Roels S, Tilmant K, Ducatelle R, 2001: p53 expression and apoptosis in melanomas of dogs and cats. Res Vet Sci, 70 (1), 19-25.
• Referans21Roels S, Tilmant K, Ducatelle R, 1999: PCNA and Ki67 proliferation markers as criteria for prediction of clinical behaviour of melanocytic tumours in cats and dogs. J Comp Pathol, 121 (1), 13-24.
• Referans22Roels S, Tilmant K, Van Daele A, Van Marck E, Ducatelle R, 2000: Proliferation, DNA ploidy, p53 overexpression and nuclear DNA fragmentation in six equine melanocytic tumours. J Vet Med A Physiol Pathol Clin Med, 47 (7), 439-448.
• Referans23Sabattini S, Renzi A, Albanese F, Fantinati M, Rigillo A, Abramo F, Tornago R, Tortorella G, Massaro M, Pagano TB, Buchholz J, Bettini G, 2018: Evaluation of Ki-67 expression in feline non-ocular melanocytic tumours. BMC Vet Res, 14 (1), 309.
• Referans24Schmid F, Brodesser D, Reifinger M, Forte S, Semp P, Eberspächer-Schweda MC, Wolschek M, Brandt S, Kleiter M, Pratscher B, 2019: Canine oral primary melanoma cells exhibit shift to mesenchymal phenotype and phagocytic behaviour. Vet Comp Oncol, 17 (3), 211-220.
• Referans25Sharma S, Chaudhary RN, Singh K, 2010: Melanoma in a Hariana cow. Haryana Vet, 49, 78.
• Referans26Sivadas CG, Nair MK, Rajan A, Ramachandran KM, 1971: Congenital melanoma in a calf. A review and case report. Br Vet J, 127 (6), 289-293.
• Referans27Smith SH, Goldschmidt MH, Mcmanus PM, 2002: A comparative review of melanocytic neoplasms. Vet Pathol, 39 (6), 651-678.
• Referans28Vadalia JV, Fefar DT, Patel PB, 2016: Surgical management of malignant melanoma in Kankrej cow. Intas Polivet, 17 (1), 98-99.
• Referans29Vijayakumar S, Lakkawar AW, Kumar R, Alphonse RMD, Nair MG, 2020: Pathomorphological studies on mesenchymal and melanocytic neoplasms of cattle. VMPH, 1 (3), 102-107.