Endoplazmik retikulum-aracılı protein yıkım yolağında görev yapan p97/VCP ve Ufd1-Npl4 adlı proteinlerin NF-?B yolağına etkisinin prostat kanser hücre hattında incelenmesi
Küçük Resim Yok
Tarih
2018
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Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Amaç: Bu çalışmada, endoplazmik retikulum-aracılı yıkım yolağındaki retrotranslokasyon basamağının anahtar proteini olan p97/VCP ve etkileşim partnerleri olan Ufd1 ve Npl4 proteinlerinin NF-?B yolağı üzerine etkileri prostat kanser hücre hattında incelendi. Gereç ve Yöntem: p97/VCP, Ufd1 ve Npl4 ifadeleri RNA interferans RNAi (RNA interferans) teknolojisi ile LNCAP hücrelerinde susturuldu ve NF-?B aktivitesi ikili lusiferaz yöntemi ile NF-?B yolağı proteinlerinin ifadesi ise immünoblotlama ile değerlendirildi. Bulgular: p97/VCP, Ufd1 ve Npl4 ifadelerinin susturulması LNCaP hücrelerinde NF-?B aktivitesini anlamlı olarak azaltmış ve I?B? protein seviyesini arttırırken fosforile NF-?B ve fosforile I?B? seviyelerini azaltmıştır. Sonuç: Retrotranslokasyon kompleks üyelerinin susturulması ile LNCaP hücrelerinde NF-?B aktivitesinin azalmasının NF-?B inhibitörü olan I?B? seviyesinin artmasına bağlı olduğu düşünülmektedir. Bulgularımız p97/VCP ve etkileşim partnerleri Ufd1-Npl4 proteinlerinin prostat kanserinde NF-?B yolağının düzenlenmesinde önemli bir etken olabileceğini önermektedir.
Aim: In this study, the effects of p97/VCP and its interacting partners Ufd1 and Npl4 proteins, which function at the retrotranslocation step of endoplasmic reticulum-associated degradation were investigated on the NF-?B pathway in the prostate cancer cell line. Materials and Methods: The expressions of p97/VCP, Ufd1 and Npl4 were silenced in LNCaP cells using RNAi (RNA interference) technology. NF-?B activity was evaluated by dual luciferase assay system and the expression of NF-?B pathway proteins were investigated using immunoblotting. Results: Silencing of expression of p97/VCP, Ufd1 and Npl4 significantly diminished NF-?B activity and increased the level of I?B? protein while decreased phosphorylated NF-?B and phosphorylated I?B? levels in LNCaP cells. Conclusion: The decrease NF-?B activity upon silencing the expressions of retrotranslocation complex members in LNCaP cells might be associated with the increased level of NF-?B inhibitor I?B?. Our data suggests that p97/VCP and its interacting partners might be important factors on the regulation of NF-?B pathway in the prostate cancer.
Aim: In this study, the effects of p97/VCP and its interacting partners Ufd1 and Npl4 proteins, which function at the retrotranslocation step of endoplasmic reticulum-associated degradation were investigated on the NF-?B pathway in the prostate cancer cell line. Materials and Methods: The expressions of p97/VCP, Ufd1 and Npl4 were silenced in LNCaP cells using RNAi (RNA interference) technology. NF-?B activity was evaluated by dual luciferase assay system and the expression of NF-?B pathway proteins were investigated using immunoblotting. Results: Silencing of expression of p97/VCP, Ufd1 and Npl4 significantly diminished NF-?B activity and increased the level of I?B? protein while decreased phosphorylated NF-?B and phosphorylated I?B? levels in LNCaP cells. Conclusion: The decrease NF-?B activity upon silencing the expressions of retrotranslocation complex members in LNCaP cells might be associated with the increased level of NF-?B inhibitor I?B?. Our data suggests that p97/VCP and its interacting partners might be important factors on the regulation of NF-?B pathway in the prostate cancer.
Açıklama
Anahtar Kelimeler
Cerrahi
Kaynak
Ege Tıp Dergisi
WoS Q Değeri
Scopus Q Değeri
Cilt
57
Sayı
3
