Farklı Tipteki Köpek Meme Karsinomlarında PCNA ve MMP-9 Ekspresyonlarının İmmunohistokimyasal Olarak Değerlendirilmesi

Öz

Bu çalışmanın amacı, köpeklerde görülen çeşitli meme adenokarsinomu alt tiplerinde hücre proliferasyonu ile invazyon-metastaz kapasitesi arasında anlamlı bir ilişki olup olmadığını PCNA ve MMP-9 gibi immunohistokimyasal belirteçler aracılığıyla ortaya koymaktır. Bu çalışmanın materyali, 2012-2020 yılları arasında anabilim dalımıza getirilen yaş ortalaması 8,4 yıl olan 6 Kangal ve 4 Setter köpeğinden alınan meme kanseri doku örnekleri oluşturdu. Köpeklerden alınan doku örnekleri,% 10 tamponlu formaldehit solüsyonunda tespit edildi. Rutin doku prosedürlerinden sonra Hematoksilen ve Eosin boyama için hazırlanan parafin bloklardan 5 μm kalınlığında kesitler alındı. Kesitler, kanser paternlerini belirlemek için ışık mikroskobu altında incelendi. İmmunohistokimyasal yöntem olarak Avidin-Biotin Peroxidase yöntemi kullanıldı. 10 meme karsinomu vakasından 4'ü intraduktal papiller karsinom, 4'ü miks karsinom, 1'i solid karsinom ve 1'i tübüler karsinom olarak sınıflandırıldı. Solid alt tipte en yüksek ortalama PCNA ve MMP-9 pozitif hücre sayısını bulduk. Sonuç olarak sadece solid alt tipte hücre proliferasyonu ile invazyon ve metastaz kapasitesi arasında pozitif bir ilişki olabileceğini düşündük.

Anahtar Kelimeler

• karsinom
• köpek
• meme
• pcna
• mmp-9

Immunohistochemically Evaluation of PCNA and MMP-9 Expressions in Different Types of Canine Mammary Carcinomas

Abstract

The aim of this study was to evaluate reveal whether there is a significant relationship between cell proliferation and invasion-metastasis capacity in various mammary adenocarcinoma subtypes observed in dogs, by means of immunohistochemical markers such as PCNA and MMP-9. The material of this study was composed of tissue samples of mammary carcinomas taken from 6 Kangal and 4 Setter dogs with an average age of 8.4 years brought to our department between 2012-2020 years. Tissue samples from dogs were fixed in 10 % buffered formaldehyde solution. After routine tissue procedures, 5 μm thick sections were taken from paraffin blocks prepared for Hematoxylin and Eosin staining. Sections were examined under light microscope to determine the cancer patterns. Avidin-Biotin Peroxidase method was used as immunohistochemical method. 4 of 10 mammary carcinoma cases were classified as intraductal papillary carcinoma, 4 as carcinoma mixed, 1 as solid carcinoma and 1 as tubular carcinoma. We found the highest mean number of PCNA and MMP-9 positive cells in solid subtype. In conclusion, we thought that a positive relationship between cell proliferation and invasion and metastasis capacity under the solid subtype.

Keywords

• carcinoma
• dog
• mammary
• pcna
• mmp-9

References

1. Referans1 Aydogan A, Ozmen O, Haligur M, Sipahi C, Ileri D, Haligur A. (2018). Immunohistochemical Evaluation of Bcl-2, ER-alpha, Caspase-3,-8,-9, PCNA and Ki-67 Expressions in Canine Mammary Carcinomas. Biotech Histochem. 93(4): 286-292.
2. Referans2 Dong H, Diao H, Zhao Y, Xu H, Pei S, Gao J, Wang J, Hussain T, Zhao D, Zhou X, Lin D. (2019). Overexpression of Matrix Metalloproteinase-9 in Breast Cancer Cell Lines Remarkably Increases the Cell Malignancy Largely via Activation of Transforming Growth Factor Beta/SMAD Signalling. Cell Prolif. 52(5): e12633.
3. Referans3 Kaszak I, Ruszczak A, Kanafa S, Kacprzak K, Król M, Jurka P. (2018). Current Biomarkers of Canine Mammary Tumors. Acta Vet Scand. 60(1): 60-66.
4. Referans4 Valencakova-Agyagosova A, Frischova Z, Sevcikova Z, Hajurka J, Lepej J, Szakallova I, Kredatusova G, Nagy V, Ledecky V. (2014). Determination of Carcinoembryonic Antigen and Cancer Antigen (CA 15-3) in Bitches with Tumours on Mammary Gland: Preliminary Report. Vet Comp Oncol. 12(3): 205-214.
5. Referans5 Vinothini G, Balachandran C, Nagini S. (2009). Evaluation of Molecular Markers in Canine Mammary Tumors: Correlation with Histological Grading. Oncol Res. 18(5-6): 193-201.
6. Referans6 Dolka I, Król M, Sapierzyński R. (2016). Evaluation of Apoptosis-Associated Protein (Bcl-2, Bax, cleaved caspase-3 and p53) Expression in Canine Mammary Tumors: An Immunohistochemical and Prognostic Study. Res Vet Sci. 105: 124-133.
7. Referans7 Kumaraguruparan R, Prathiba D, Nagini S. (2006). Of Humans and Canines: Immunohistochemical Analysis of PCNA, Bcl-2, p53, Cytokeratin and ER in Mammary Tumours. Res Vet Sci. 81(2): 218-224.
8. Referans8 Muhammadnejad A, Keyhani E, Mortazavi P, Behjati F, Haghdoost IS. (2012). Overexpression of Her-2/Neu in Malignant Mammary Tumors; Translation of Clinicopathological Features from Dog to Human. Asian Pac J Cancer Prev. 13(12): 6415-6421.

9. Referans9 Senhorello ILS, Terra EM, Sueiro FAR, Firmo BF, Anai LA, Goloni C, Canavari IC, Ampuero RAN, Pereira RS, Tinucci-Costa M. (2020). Clinical Value of Carcinoembryonic Antigen in Mammary Neoplasms of Bitches. Vet Comp Oncol. 18 (7069): 315–323.
10. Referans10 Ranganath GJ, Kumar Ram, Vishwanatha Reddy AP, Mayilkumar K, Pawaiya RVS, Maiti SK. (2011). Comparative Study on the Expression Pattern of the Proliferating Cell Markers PCNA and Ki67 in Canine Mammary Tumours. Ind J Vet Pathol. 35(1): 13-17.
11. Referans11 Chen YC, Chang SC, Huang YH, Lee YJ, Chang CC, Liao JW, Hsu WL. (2019). Expression and the Molecular Forms of Neutrophil Gelatinase-associated Lipocalin and Matrix Metalloproteinase 9 in Canine Mammary Tumours. Vet Comp Oncol. 17(3): 427-438.
12. Referans12 Veena P, Suresh Kumar RV, Raghavender KBP, Srilatha Ch, Rao TSC. (2014). Immunohistochemical Detection of p53 in Canine Mammary Tumors. Indian J Anim Res. 48(2): 204-206.
13. Referans13 Klopfleisch R, Klose P, Weise C, Bondzio A, Multhaup G, Einspainer R, Gruber AD. (2010). Proteome of Metastatic Canine Mammary Carcinomas: Similarities to and Differences from Human Breast Cancer. J Proteome Res. 9(12): 6380-6391.
14. Referans14 Carvalho MI, Pires I, Prada J, Lobo L, Queiroga FL. (2016). Ki-67 and PCNA Expression in Canine Mammary Tumors and Adjacent Nonneoplastic Mammary Glands: Prognostic Impact by a Multivariate Survival Analysis. Vet Pathol. 53(6): 1138-1146.
15. Referans15 Munday JS, Ariyarathna H, Aberdein D, Thomson NA. (2019). Immunostaining for p53 and p16CDKN2A Protein is not Predictive of Prognosis for Dogs with Malignant Mammary Gland Neoplasms. Vet Sci. 6(1): 34.
16. Referans16 Goldschmidt MH, Peña L, Zappulli V. (2017). Tumors of the Genital Systems. In : Tumors of Domestic Animals. Moulton JE (ed), 5th ed. pp. 723-764, Wiley Blackwell, Iowa, USA.
17. Referans17 Łopuszyński W, Hellmén E. (2015). Cell Proliferation Study in Canine Mammary Carcinomas. Int J Vet Health Sci Res. 3(2): 39-45.
18. Referans18. Preziosi R, Sarli G, Benazzi C, Marcato PS. (1995). Detection of Proliferating Cell Nuclear Antigen (PCNA) in Canine and Feline Mammary Tumours. J Comp Path. 113(4): 301-313.
19. Referans19 Zuccari DAPC, Pavam MV, Terzian ACB, Pereira RS, Ruiz CM, Andrade JC. (2008). Immunohistochemical Evaluation of E-cadherin, Ki-67 and PCNA in Canine Mammary Neoplasias: Correlation of Prognostic Factors and Clinical Outcome. Pesq Vet Bras. 28(4): 207-215.
20. Referans20 Funakoshi Y, Nakayama H, Uetsuka K, Nishimura R, Sasaki N, Doi K. (2000). Cellular Proliferative and Telomerase Activity in Canine Mammary Gland Tumors. Vet Pathol. 37(2): 177-183.
21. Referans21 Peña LL, Nieto AI, Pérez-Alenza D, Cuesta P, Castaño M. (1998). Immunohistochemical Detection of Ki-67 and PCNA in Canine Mammary Tumors: Relationship to Clinical and Pathologic Variables. J Vet Diagn Invest. 10(3): 237-246.
22. Referans22 Zacchetti A, van Garderen E, Teske E, Nederbragt H, Dierendonck JH, Rutteman GR. (2003). Validation of the Use of Proliferation Markers in Canine Neoplastic and Non-Neoplastic Tissues: Comparison of KI-67 and Proliferating Cell Nuclear Antigen (PCNA) Expression Versus in Vivo Bromodeoxyuridine Labelling by Immunohistochemistry. APMIS. 111(3): 430-438.
23. Referans23 İlhan F, Metin N, Türkütanıt Birincioğlu S. (2008). Immunohistochemical Detection of PCNA and p53 in Mammary Tumours and Normal Tissues in Dogs. Revue Méd Vét. 159(5): 298-304.
24. Referans24 Silva FB, Leite Jda S, de Mello MF, Ferreira AM (2016). Validation of a Low-cost Modified Technique for Constructing Tissue Microarrays For Canine Mammary Tumor Analysis. Pathol Res Pract. 212(9): 783-790.
25. Referans25 Lokesh JV, Kurade NP, Shivakumar MU, Sharma AK, Maiti SK. (2014). Evaluation of BcL-2 and PCNA Expression and Mitotic Index in Spontaneous Canine Tumors. Adv Anim Vet Sci. 2(1): 63-66.
26. Referans26 Hirayama K, Yokota H, Onai R, et al. (2002). Detection of Matrix Metalloproteinases in Canine Mammary Tumours: Analysis by Immunohistochemistry and Zymography. J Comp Pathol. 127(4): 249-256.
27. Referans27 Aresu L, Giantin M, Morello E, et al. (2011). Matrix Metalloproteinases and Their Inhibitors in Canine Mammary Tumors. BMC Vet Res. 7: 33.
28. Referans28 Nowak M, Madej JA, Podhorska-Okolow M, Dziegiel P. (2008). Expression of Extracellular Matrix Metalloproteinase (MMP-9), E-cadherin and Proliferation-associated Antigen Ki-67 and Their Reciprocal Correlation in Canine Mammary Adenocarcinomas. In Vivo. 22(4): 463-469.
29. Referans29 Santos AA, Lopes CC, Marques RM, Amorim IF, Gärtner MF, de Matos AJ. (2012). Matrix Metalloproteinase-9 Expression in Mammary Gland Tumors in Dogs and its Relationship with Prognostic Factors and Patient Outcome. Am J Vet Res. 73(5): 689-697.
30. Referans30 Raposo TP, Beirão BC, Pires I, et al. (2016). Immunohistochemical Expression of CCR2, CSF1R and MMP9 in Canine Inflammatory Mammary Carcinomas. Anticancer Res. 36(4): 1805-1813.