Hydrated C60 fullerene, glioblastoma hücre hattında parthanatosu arttırır ve otofaji ile ilgili biyobelirteçleri indükler

Öz

Glioblastoma, kemo ve radyoterapiye karşı dirençli, en agresif beyin kanseri tiplerinden biridir. C60 fulleren türevi nanopartiküller, çeşitli modellerde antikanser aktivite amacı ile geliştirilmektedir. Birçok kemoterapi ajanının aksine, bu fulleren çeşitli konsantrasyonlarda toksik değildir. C60 fulleren, birçok biyomedikal uygulama için umut verici bir adaydır. Bu nedenle, suda çözünür hydrated C60 fullerene'in (HyC60Fn) insan glioblastoma U373 hücresinde PARP, Beclin1, LC3 ve GFAP ekspresyonu üzerindeki etkileri araştırılmıştır. Hücre canlılığı ve göçü, sırasıyla MTT ve yara iyileşmesi testi ile belirlendi. PARP, Beclin1 ve LC3 ekspresyonu western blot ile ve GFAP ise immünositokimya ile tespit edildi. 0.5 – 2.0 µM doz aralığındaki HyC60Fn, doza bağlı bir şekilde hücre canlılığını azalttığı belirlendi. Ayrıca, HyC60Fn 1.0 ve 2.0 µM dozları, glioblastoma hücre göçünü belirgin şekilde bastırmıştır. Mekanizma olarak, HyC60Fn'nin otofaji belirteçleri olarak Beclin-1'i ve LC3-II/LC3-I ekspresyon oranını belirgin şekilde yukarı regüle ettiği belirlendi. Ayrıca, suda çözünür HyC60Fn’nin PARP fragmanı ve bu durumun doğal sonuç olarak glioblastoma U373 hücrelerinde parthanatos aktive ettiği belirlendi. Mevcut sonuçlar, HyC60Fn'nin, glioblastoma hücrelerinde şiddetli otofaji akışı ve parthanatos kombinasyonu yoluyla anti-tümör etkisini başlatabildiğini göstermektedir. Bu nedenle HyC60Fn, glioblastoma hücrelerinin reaktivitesini ve programlanmış hücre ölümünü modüle ederek en azından kısmen hücre ölüm mekanizmasını etkiler. Bulgularımız, HyC60Fn 'in umut verici bir kanser karşıtı terapötik olabileceğini ve bu konuda daha fazla çalışmanın gerekli olduğunu göstermektedir.

Anahtar Kelimeler

• Glioblastoma
• C60 hydrated fullerene
• Otofaji
• Parthanatos

Hydrated C60 fullerene enhances parthanatos and induces autophagy-related biomarkers in glioblastoma cell line

Abstract

Glioblastoma is one the most aggressive type of brain cancers, which is resistant to resistant chemo- and radio-therapy. Nanoparticles of C60 fullerene derivates develop anticancer activity in various models. In contrast to many chemotherapy agents, this fullerene absolutely nontoxic in wide range of concentrations with respect to normal cells. C60 fullerene is a promising candidate for many biomedical applications. Therefore, we investigated the effect of water soluble hydrated C60 fullerene (HyC60Fn) on the expression of PARP, Beclin1, LC3, and GFAP in human glioblastoma U373 cell. Cell viability and migration were detected by MTT and wound healing-scratch assay, respectively. The expression of PARP, Beclin1, and LC3 were analyzed by western blotting and GFAP was detected with immunocytochemistry. HyC60Fn in a range of doses 0.5 – 2.0 µM decreased cell viability in a dose-dependent manner. Furthermore, the doses of HyC60Fn 1.0 and 2.0 µM noticeably suppressed glioblastoma cell migration. Mechanistically, we defined that HyC60Fn markedly up-regulated Beclin-1 and ratio of LC3-II/LC3-I expression as autophagy markers. Furthermore, water soluble HyC60Fn activated cleaved PARP fragment and consequently parthanatos in glioblastoma U373 cells. Present results demonstrate that HyC60Fn could initiate anti-tumor effect via the combination of severe autophagy flux and parthanatos in glioblastoma cells. Thus, HyC60Fn affects the cell death machinery, at least partially, through modulating glioblastoma cells reactivity and programmed cell death. Our findings suggest that pristine hydrated C60 fullerene could be a promising anti-cancer therapeutics and further study is required

Keywords

• Glioblastoma
• C60 hydrated fullerene
• Autophagy
• Parthanatos

Kaynakça

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