The Investigation of Protocadherin Gene Family Members PCDH8, PCDH10, and PCDH17 as Epigenetic Biomarkers in Blood Samples of Patients with Bladder Tumor

Abstract

Objective: The aim of the study was to evaluate the potential tumor-specific effects of peripheral blood DNA methylation profiles of protocadherin 8 (PCDH8), protocadherin 10 (PCDH10), and protocadherin 17 (PCDH17) genes who belong to the protocadherin gene family, in patients with bladder tumors and healthy controls. The potential association between methylation profiles of the investigated three genes and demographic and/or clinicopathologic data was a second target of the study.

Material and Methods: Genomic DNA extracted from peripheral blood samples of patients with bladder carcinoma (n=80; low-grade: 40, high-grade: 40) and healthy controls (n=40) was subjected to bisulphite modification and methylation-specific PCR for hypermethylation analyses of promoter regions.

Results: PCDH8 methylation profile was observed to be increased with age. Though PCDH10 and PCDH17 methylation patterns did not reflect a difference in patients since partial methylation profiles were also almost completely detected in controls; PCDH8 displayed a more different blood epigenetic profile.

Conclusion: The association of blood methylation pattern of PCDH8 with aging process was shown in our study. We recommend the investigation of the status of PCDH8 methylation in blood in larger cohorts and with more sensitive methods such as MethyLight to draw a more precise conclusion in terms of its effect in bladder carcinoma.

Keywords

• Bladder cancer
• DNA methylation
• peripheral blood
• PCDH8,
• PCDH10
• PCDH17

MESANE TÜMÖRLÜ HASTALARIN KANLARINDA EPİGENETİK BİYOBELİRTEÇLER OLARAK PROTOKADERİN GEN AİLESİ ÜYELERİ PCDH8, PCDH10 VE PCDH17’NİN ARAŞTIRILMASI

Abstract

Amaç: Çalışmanın amacı mesane tümörlü hastalarda ve sağlıklı kontrollerde protokaderin gen ailesine ait olan protokaderin 8 (PCDH8), protokaderin 10 (PCDH10) ve protokaderin 17 (PCDH17) genlerinin periferal kan DNA metilasyon profillerini analiz ederek bu profillerin tümör-spesifik olası etkilerini değerlendirmektir. Araştırılan üç genin metilasyon profiliyle spesifik demografik ve/veya klinikopatolojik veriler arasındaki olası ilişkinin araştırılması da çalışmanın ikinci hedefidir.

Gereç ve Yöntemler: Mesane karsinomalı (n=80; düşük dereceli: 40, yüksek dereceli: 40) ve sağlıklı kontrollerin (n=40) periferal kan örneklerinden ekstrakte edilen genomik DNA promotor bölgelerinin hipermetilasyon analizi için bisülfit modifikasyon ve metilasyon-spesifik PCR işlemine maruz bırakılmıştır.

Bulgular: PCDH8 metilasyon profilinin yaşla birlikte artış gösterdiği gözlenmiştir. Neredeyse kontrollerin tamamında da bu genler bakımından kısmi metilasyon profilleri belirlendiği için hasta örneklerindeki PCDH10 ve PCDH17 metilasyon paternleri bir farklılık oluşturmamakla beraber PCDH8 daha farklı bir kan epigenetik profili sergilemiştir.

Sonuç: PCDH8 geninin kandaki metilasyon modelinin yaşlanma süreciyle ilişkisi çalışmamızda gösterilmiştir. Mesane kanserindeki etkisi açısından daha kesin bir sonuca varabilmek için PCDH8’in kandaki metilasyon durumunun daha yüksek sayılı kohortta ve MethyLight gibi daha duyarlı metotlarla araştırılmasını tavsiye etmekteyiz.

Keywords

• Mesane kanseri
• DNA metilasyonu
• periferik kan
• PCDH8
• PCDH10
• PCDH17

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