Dosetaksel Dirençli Prostat Kanseri Hücrelerinde Timokinon Tarafından Otofajik Hücre Ölümünün İndüklenmesi

Amaç: Prostat kanserinde (prostate cancer, PCa) edinilen dosetaksel (docetaxel, DOC) direnci hala klinik bir sorundur. Kemoterapi tedavisinde başarısızlığa ek olarak tümör nüksüne neden olmaktadır. Bu nedenle, DOC’a dirençli PCa tedavisinde yeni ve daha etkili bileşiklere ihtiyaç duyulmaktadır. Bu çalışmanın amacı, Nigella sativa L. bitkisinin etken bileşenlerinden biri olan timokinon (thymoquinone, TQ)’un, DOC dirençli prostat kanseri hücreleri üzerindeki olası sitotoksik ve hücre ölümünü tetikleyici aktivitelerinin araştırılmasıdır. Gereç ve Yöntemler: DOC dirençli PC3 hücreleri (DOC-R/PC3), büyüme ve bölünme yeteneklerini geliştirene kadar artan DOC (1-10 nM) konsantrasyonlarında devamlı kültürle çoğaltıldı. Hücre canlılığı, MTT yöntemi kullanılarak belirlendi. MuseTM Annexin V & Dead Cell kiti, apoptotik hücre ölümünün tespiti için kullanıldı. Otofajik vakuoller spesifik boya kullanılarak gösterildi. TQ mualemesi sonucu LC3-I, LC3-II ve Beclin-1 protein düzeylerindeki değişiklikler western blot analizi ile araştırıldı. Bulgular: TQ muamelesi, DOC-R/PC3 hücrelerinin canlılığını doza ve zamana bağlı olarak inhibe etti (p=0.014). DOC-R/PC3 için TQ'nun IC50 değeri 72. saatte 60 µM olarak hesaplandı. TQ uygulaması, DOC dirençli prostat kanseri hücrelerinde apoptotik hücre ölümünü indüklemedi, ancak otofajik vakuol oluşumunu indükledi. Ayrıca TQ ile muamele edilen DOC-R/PC3 hücrelerinde Beclin-1 ve LC3-II protein seviyelerinin arttığı, ancak DOC-R/PC3 hücrelerinde LC3-I seviyelerinin azaldığı tespit edildi. Sonuç: Tüm bu sonuçlar, TQ'nun gelecekte DOC dirençli prostat kanseri için yeni bir terapötik ajan olabileceğini göstermektedir.

Anahtar Kelimeler:

otofaji, Beclin-1, LC3, prostat kanseri, direnç, timokinon

Induction of Autophagic Cell Death by Thymoquinone in Docetaxel Resistant Prostate Cancer Cells

Aim: Acquired docetaxel (DOC) resistance of prostate cancer (PCa) is still a clinical problem. In addition to failure in chemotherapy treatment, it causes tumor recurrence. Therefore, novel and more effective compounds are needed in DOC-resistant PCa treatment. This study aimed to investigate the possible cytotoxic and cell death-inducing activities of thymoquinone (TQ), one of the main active components of Nigella sativa L., on DOC-resistant prostate cancer cells. Material and Methods: DOC-resistant PC3 cells (DOC-R/PC3) were developed by the continuous culture with increment concentrations of DOC (1-10 nM) until they improved their growth and division abilities. The cell viability was determined by MTT assay. The MuseTM Annexin V & Dead Cell kit was performed to detect apoptotic cell death. Autophagic vacuoles were observed by staining autophagic vacuoles. The levels of LC3I, LC3II and Beclin-1 proteins were investigated via western blot analysis. Results: TQ inhibited the viability of DOC-R/PC3 cells in a dose- and time-dependent manner (p=0.014). The IC50 value of TQ for DOC-R/PC3 cells was calculated as 60 µM at 72 h. Treatment of TQ did not induce apoptotic cell death in DOC-resistant prostate cancer cells but induced the formation of autophagic vacuoles. Moreover, Beclin-1 and LC3-II protein levels were increased in TQ-treated DOC-R/PC3 cells, however, LC3-I levels were decreased in DOC-R/PC3 cells. Conclusion: All these results show that TQ may become a new therapeutic target for DOC-resistant prostate cancer in the future.

Keywords:

autophagy, Beclin-1, LC3, prostate cancer, resistance, thymoquinone,

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