Kedilerin Yassı Hücreli Karsinomlarında İntermedier Filamentler, p53 Geni, Hücre Proliferasyonu, Metastaz ve Apoptozis

Abstract

Bu çalışmada, kedi YHK'larının hücresel kökenini ortaya çıkarmak için intermedier filamentler, Pan Sitokeratin (Pan SK), Vimentin, Desmin ve S-100 değerlendirilmiştir. Tümör mikroçevresindeki kanserle ilişkili fibroblastlar (KAF'lar) için Alfa-Düz Kas Aktini (α-SMA) ve bir tümör baskılayıcı gen olan p53 araştırıldı. Prolifere Hücre Nükleus Antijeni (PCNA) ekspresyonu, hücre proliferasyon indeksi için değerlendirildi. Matriks Metalloproteinaz-9 (MMP-9) immünoreaktivitesi, metastaz ve istila kapasitesi açısından değerlendirildi. Ayrıca apoptoz mekanizması için proapoptotik Bax geni, antiapoptotik Bcl-2 geni, kaspaza bağımlı yol olan Kaspaz-3 ve kaspazdan bağımsız yol olan Apoptoz İndükleyici Faktör (AIF) ekspresyonlarının ortaya çıkarılması amaçlanmaktadır. Bu çalışmada Patoloji Anabilim Dalı'na rutin histopatolojik inceleme için getirilen 7 kediden alınan biyopsi örnekleri kullanıldı. Tümör dokusu örnekleri %10'luk formaldehit solüsyonunda fikse edildi. Rutin doku takip işlemlerinden sonra hazırlanan parafin bloklardan 5 µm kalınlığında seri kesitler alındı. Kesitler üzerinde Hematoksilen & Eozin (H&E) boyaması yapıldı. İmmünohistokimyasal boyama olarak avidin-Biotin Peroksidaz-Tekniği (ABC) kullanıldı. Tümörlerin epitelyal-mezenkimal geçişe sahip olduğu, çok yüksek proliferasyon indeksi sergilediği, p53 mutasyonuna sahip olduğu ve düşük metastaz/yüksek invazyon kapasitesi gösterdiği belirlendi. Bax/Bcl-2 oranının proapoptotik Bax lehine arttığı ve kaspazdan bağımsız apoptozun kaspaz bağımlı apoptozdan daha baskın olduğu ortaya çıktı.

Keywords

• apoptoz
• hücresel proliferasyon
• intermedier filamentler
• kedilerin yassı hücreli karsinomu
• metastaz

Intermediate Filaments, P53 Gene, Cellular Proliferation, Metastasis and Apoptosis in Feline Squamous Cell Carcinomas

Abstract

In this study, intermediate filaments, Pan Cytokeratin (Pan CK), Vimentin, Desmin and S-100 were evaluated to reveal the cellular origin of feline SCCs. Alpha-Smooth Muscle Actin (α-SMA) for cancer-related fibroblasts (CAFs) in the tumor microenvironment, and p53, a tumor suppressor gene, were investigated. Proliferating Cell Nuclear Antigen (PCNA) expression was evaluated for the cell proliferation index. Matrix Metalloproteinase-9 (MMP-9) immunoreactivity was evaluated for the metastasis and invasion capacity. In addition, it is aimed to reveal the expressions of proapoptotic Bax gene, antiapoptotic Bcl-2 gene, caspase-dependent pathway Caspase-3 and caspase-independent pathway Apoptosis Inducing Factor (AIF) for apoptosis mechanism. Biopsy samples taken from 7 cats brought to Department of Pathology for routine histopathological examination were used in this study. Tumor tissue samples were fixed in 10% formaldehyde solution. Serial sections of 5 μm thickness were taken from the paraffin blocks prepared after routine tissue follow-up procedures. Hematoxylin & Eosin (H&E) staining was performed on the sections. Avidin-Biotin Peroxidase-Technique (ABC) was used as immunohistochemical staining. It was determined that the tumors had epithelial-mesenchymal transition, exhibited a very high proliferation index, had p53 mutation, and showed low metastasis/high invasion capacity. It was revealed that Bax/Bcl-2 ratio increased in favor of proapoptotic Bax, and caspase-independent apoptosis was more dominant than caspase-dependent apoptosis.

Keywords

• Apoptosis
• cellular proliferation
• feline squamous cell carcinoma
• intermediate filaments
• metastasis

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