The stimulation of apoptosis via mitochondrial (intrinsic) pathway in HeLa cells by some anthraquinone derivates

Abstract

Objective: The aim of this study was to investigate the effects of Ant-3, -4, and -6 anthraquinone derivatives on the mitochondrial (intrinsic) apoptotic pathway in HeLa cells. Methods: Mitochondrial membrane potential changes, Caspase-9 activity, apoptotic activity, and protein expression levels involved in the intrinsic pathway were measured. The TUNEL method was used to determine the apoptotic index. Results: Maximum Caspase-9 activation was observed after 6 hours of incubation with Ant-6 (3 μM) and Ant-4 (5 μM).The highest mitochondrial membrane potential change occurred after 36 hours of treatment with Ant-4 (5 μM).The apoptotic index, determined by the TUNEL method, peaked in the Ant-4 (5 μM) group after 48 hours. Increased expression of AIF and EndoG proteins was observed after 48 hours in HeLa cells treated with Ant-4 and mitoxantrone. Conclusion: These findings show promising results in in vitro experiments. However, further clinical studies are needed to validate the therapeutic potential of these newly synthesized anthraquinone derivatives in the treatment of cervical cancer.

Keywords

• Anthraquinone
• apoptosis
• caspase-9
• HeLa
• intrinsic pathway
• Western Blot

Bazı Antrakinon Türevleri ile HeLa Hücrelerinde Mitokondriyal (İntrinsik) Yolak Üzerinden Apoptozun Uyarılması

Abstract

Amaç: Bu çalışmanın amacı, Ant-3, -4 ve -6 antrakinon türevlerinin HeLa hücrelerinde mitokondriyal (intrinsik) apoptotik yolak üzerindeki etkilerini araştırmaktır. Yöntemler: Mitokondriyal membran potansiyeli değişiklikleri, Kaspaz-9 aktivitesi, apoptotik aktivite ve intrinsik yolağa dahil olan proteinlerin ekspresyon seviyeleri ölçülmüştür. Apoptotik indeksin belirlenmesinde TUNEL yöntemi kullanılmıştır. Bulgular: Ant-6 (3 μM) ve Ant-4 (5 μM) ile 6 saatlik inkübasyon sonrası maksimum Kaspaz-9 aktivasyonu gözlenmiştir. Mitokondriyal membran potansiyelindeki en yüksek değişiklik, Ant-4 (5 μM) ile 36 saatlik uygulama sonrası meydana gelmiştir. TUNEL yöntemiyle belirlenen apoptotik indeks, Ant-4 (5 μM) grubunda 48 saat sonunda en yüksek seviyeye ulaşmıştır. Ant-4 ve mitoksantron ile tedavi edilen HeLa hücrelerinde 48 saat sonunda AIF ve EndoG proteinlerinin ekspresyonunda artış gözlenmiştir. Sonuç: Bu bulgular, in vitro deneylerde umut verici sonuçlar ortaya koymaktadır. Bununla birlikte, bu yeni sentezlenen antrakinon türevlerinin rahim ağzı kanseri tedavisindeki terapötik potansiyelini doğrulamak için ileri klinik çalışmalara ihtiyaç vardır.

Keywords

• Antrakinon
• apoptoz
• kaspaz-9
• HeLa
• intrinsik yolak
• Western Blot

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