Usnik Asit SH-SY5Y Nöroblastom Hücrelerinde Tümör Baskılayıcı Genleri Aktive Ederek Apoptozu Tetikler ve İnvazyonu Azaltır

Öz

Usnik asit, Usnea diffracta Vain'den kaynaklanan bir dibenzofuran bileşiğidir ve anti-inflamatuar, antifungal ve antikanser etkileri vardır. Ancak, antitümör etkilerinin moleküler mekanizması tam olarak açıklanmamıştır. Bu çalışma, usnik asidin nöroblastom hücrelerinde antikanser aktivitesinin altında yatan apoptoz ve otofaji aracılı mekanizmaları belirlemeyi amaçlamaktadır. Usnik asidin SH-SY5Y nöroblastom hücre canlılığı üzerine etkileri CCK-8 testi, hücre migrasyonu üzerine etkileri yara iyileşme deneyi, invazyon, apoptoz, otofaji ve tümör baskılayıcı genler üzerine etkileri ise Western blot ile değerlendirilmiştir. Usnik asidin SH-SY5Y nöroblastom hücre canlılığını doz ve zaman bağımlı olarak azalttığı gösterilmiş ve IC50 dozu 24 saatte 10 μM olarak belirlenmiştir. Usnik asidin nöroblastom hücre migrasyonu ve invazyonunu anlamlı olarak azalttığı gösterilmiştir. Kontrol grubu yara alanı 0. saatte ortalama 628,9 ± 35,59 cm2 iken 24 saatte 234,8 ± 30,98 cm2 olarak belirlenmiştir. Usnik asit yara alanı ise 0. saatte ortalama 895,2 ± 30,07 cm2 iken 24 saatte 801,9 ± 15,23 cm2’dir. Usnik asidin nöroblastom hücre apoptozunu indüklediği belirlenmiştir. Usnik asit kesilmiş kaspaz-3 ekspresyonunda 4 katlık bir artışa neden olmuş, otofaji markırlarında ise bir değişiklik oluşturmamıştır. Usnik asit uygulaması SH-SY5Y nöroblastom hücrelerinde tümör baskılayıcı genlerin ekspresyonunda artışa neden olmuştur. Sonuç olarak, usnik asidin nöroblastom hücrelerinde tümör baskılayıcı genleri aktive ederek apoptozu tetikleyebileceği, böylelikle invazyon, migrasyon ve hücre canlılığında azalışa neden olabileceği gösterilmiştir. Usnik asidin nöroblastom hücrelerinde antikanser aktivitesinin altında yatan mekanizmaların daha ileri çalışmalarla desteklenmesi gerekmektedir.

Anahtar Kelimeler

• Usnic asit
• nöroblastom
• apoptoz
• otofaji
• invazyon

Usnic Acid Triggers Apoptosis and Reduces Invasion by Activating Tumor Suppressor Genes in SH-SY5Y Neuroblastoma Cells.

Öz

Usnic acid is a dibenzofuran compound derived from Usnea diffracta Vain and has anti-inflammatory, antifungal and anticancer activities. However, the molecular mechanism of its antitumor effects has not been fully elucidated. This study aims to identify the apoptosis and autophagy-mediated mechanisms underlying the anticancer activity of usnic acid in neuroblastoma cells. The effects of usnic acid on SH-SY5Y neuroblastoma cell viability were evaluated by CCK-8 test, its effects on cell migration by wound healing assay, and its effects on invasion, apoptosis, autophagy and tumor suppressor genes by Western blot. Usnic acid was shown to reduce SH-SY5Y neuroblastoma cell viability in a dose and time dependent manner, and the IC50 dose was determined as 10 μM at 24 hours. Usnic acid has been shown to significantly reduce neuroblastoma cell migration and invasion. The control group wound area was determined to be 628.9 ± 35.59 cm2 on average at hour 0, while it was determined to be 234.8 ± 30.98 cm2 at 24 hours. Usnic acid wound area was 895.2 ± 30.07 cm2 at 0 hour and 801.9 ± 15.23 cm2 at 24 hours. It has been determined that usnic acid induces neuroblastoma cell apoptosis. Usnic acid caused a 4-fold increase in the expression of cleaved caspase-3, but did not cause any change in autophagy markers. Usnic acid application caused an increase in the expression of tumor suppressor genes in SH-SY5Y neuroblastoma cells. As a result, it has been shown that usnic acid can trigger apoptosis by activating tumor suppressor genes in neuroblastoma cells, thereby causing a decrease in invasion, migration and cell viability. The mechanisms underlying the anticancer activity of usnic acid in neuroblastoma cells need to be supported by further studies.

Anahtar Kelimeler

• usnic acid
• neuroblastoma
• apoptosis
• autophagy
• invasion

Kaynakça

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