Poly I:C'nin İndüklediği TLR3 Aktivasyonunun Prostat Kanseri Hücreleri Olan PC-3 (Hormona Duyarsız) ve LNCaP'ın (Hormona Duyarlı) Oksidatif Stres Düzeyine Etkileri

Öz

Amaç: Poly I:C ile indüklenen TLR3 aktivasyonunun iki farklı prostat kanseri hücresinde [PC-3 (hormona duyarsız) ve LNCaP (hormona duyarlı)] oksidatif stres üzerindeki rolünü ilk kez belirlemeyi amaçladık. Bu amaçla lipid peroksidasyonu (MDA), hidrojen peroksit (H2O2) ve prolin miktarlarına, süperoksit dismutaz (SOD) enzim aktivitesine bakılmıştır.
Yöntem: Reseptör uyarımı için gerekli olan ve hücre canlılığını destekleyen optimal Poly I:C doz ve süresi WST-1 analizi ile belirlendi. Biyokimyasal parametrelere spektrofotometrik yöntemler ile tayin edildi.
Bulgular: Poly I:C'nin PC-3 ve LNCaP hücreleri üzerinde daha az sitotoksik konsantrasyonunu olarak 5 µM belirlendi. SOD aktivitelerininde LNCaP hücrelerinde önemli bir artış 6 ve 24 saat sonra gözlenmedi. 6 saat boyunca PC-3 ve LNCaP hücrelerinin MDA seviyelerinde önemli bir artış belirlenirken, 24 saat sonra Poly I:C LNCaP hücrelerinde önemli bir düşüş gözlemlendi. LNCaP hücrelerininde H2O2 konsantrasyonunda önemli artış tespit edildi. Buna karşın 6 ve 24 saatlik Poly I:C uygulamalarından sonra PC-3 hücrelerinde H2O2 konsantrasyonunda önemli bir düşüş gözlendi. Prolin seviyesi LNCaP hücrelerinde 24 saat boyunca önemli bir artış gösterdi ancak PC-3 hücrelerinde hem 6 hem de 24 saat sonra prolin seviyesinde değişiklik olmadı.
Sonuç: Hormona duyarlı LNCaP hücrelerinde MDA, H2O2 ve SOD aktivite düzeyleri anlamlı olarak yüksek bulunurken Poly I:C ile tedavi edilen metastatik ve hormona duyarsız PC-3 hücrelerinde önemli bir değişiklik bulunmamıştır. İstatiksel veriler kontrol grubuyla karşılaştırıldığında p<0,05 ve p<0,001 düzeyinde anlamlı olarak farklıdır.

Anahtar Kelimeler

• Kanser
• prolin
• Poly I:C,
• toll benzeri reseptör 3

Poly I:C-Induced TLR3 Activation on Oxidative Stress Levels in PC-3 (Hormone-Insensitive) and LNCaP (Hormone-Sensitive) as Prostate Cancer Cells

Öz

Objective: We aimed to determine the role of Poly I:C-induced TLR3 activation on oxidative stress in two different prostate cancer cells [PC-3 (hormone insensitive) and LNCaP (hormone-sensitive)] for the first time. For this purpose, lipid peroxidation (MDA), hydrogen peroxide (H2O2) and proline amounts and superoxide dismutase (SOD) enzyme activity were examined.
Methods: The optimal concentration and time required for receptor stimulation with Poly I:C cells were determined by WST-1 analysis. Spectrophotometric methods determined biochemical parameters.
Results: The less cytotoxic concentration of 5 µM of Poly I:C on PC-3 and LNCaP cells was determined. A significant increase was observed in LNCaP cells in SOD activities after 6 and 24 hours. A significant increase in PC-3 and LNCaP cells' MDA levels was determined over 6 hours, while a significant decrease was observed in Poly I:C LNCaP after 24 hours. A significant increase in H2O2 concentration was detected in LNCaP cells, but a significant decrease was observed in PC-3 after 6 and 24 hours. The proline level showed a significant increase in LNCaP over 24 hours but not in the proline level in PC-3 cells after 6 and 24 hours.
Conclusion: The MDA, H2O2 and SOD activity levels were found to be significantly higher in hormone-sensitive LNCaP cells, while no significant changes were found in PC-3 cells treated with Poly I:C. Results were significantly different at the level of p<0.05 and p<0.001.

Anahtar Kelimeler

• Cancer
• proline
• Poly I:C
• toll-like receptor 3

Teşekkür

The authors thank Dr. Ozlem Aksoy from the Department of Biology at Kocaeli University for supporting our experiments.

Kaynakça

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