FİLİZ ÖZDEMİR, Zerrin İNCESU, Mesut ŞENA, Nur İpek ÖNDERA, Miris DİKMEN

HepG2 hücrelerinde Vinkristin sülfat/?-viniferin kombinasyonunun vinkristin sülfat ile karşılaştırıldığında artmış etkinliğinin vurgulanması

Amaç: Bu çalışma HepG2 hücre hattında geç apoptoz ve mitokondriyal yolakta ?-viniferin (?-VNF)'in etkisini incelemek için dizayn edildi. Bu amaç için, HepG2 hücre dizisinde anti-kanser ilaç olarak kullanılan vinkristin sülfat (VNC) ve ?-VNF farklı zaman aralıklarında tek başına ve kombine olarak verildi. Yöntemler: Hücrelerin mitokondriyal membrane potansiyeli (??m) katyonik boya ile, RT-PCR ile Bax, Bcl-2, kaspaz-3 aktivitesi de kit kullanılarak değerlendirildi. Apoptotik aktivite in situ TUNEL ile tespit edildi. Bulgular: 98.3µM ?-VNF, 52.5µM VNC ve 11.25+15.8µM VNC+?-VNF kombinasyonu kontrol grubu ile mukayese edildiği zaman, ??m değişimleri 6. saatte sırasıyla %19,5, %5,5, %24,6 ve %3,5 olarak bulundu. Bu sonuçlar 6. Saatte kombinasyon grubunun (%24,6) erken apoptoza gittiğini göstermiştir. Bax mRNA ekspresyonu 24. saatte VNC+?-VNF grubu kontrol grubu ile mukayese edildiğinde (%160), kaspaz-3 aktivasyonu VNC+?-VNF (11.25+15.8 µM) grubu kontrol ile mukayese edildiğinde 48. saatte artış göstermiştir. 24 saat içerisinde, HepG2 hücrelerinde DNA kırıklarının tespit edilmiş olması doğrudan apoptozis ile ilişkili oldugunu göstermektedir. Sonuç: Bu bulgular apoptozise yol açan VNC+ ?-VNF grubunun, tek başına kullanılan ajanın (VNC) oluşturacağı muhtemel toksik dozu azaltabileceğini göstermektedir.

Implications of enhanced effectiveness of vincristine sulfate/?-viniferin combination compared to vincristine sulfate only on HepG2 cells

Objective: This study was designed to investigate the effects of ?-viniferin (?-VNF) on the mitochondrial pathway of apoptosis and on late apoptosis in HepG2 cell lines. To observe these effects, ?-VNF and vincristine sulfate (VNC), anti-cancer drugs used for treatment on HepG2 cells, were administered either alone or in combination at different time intervals. Methods: Mitochondrial membrane potential changes in the cells (??m) were evaluated using cationic dye JC-1, while Bax, Bcl2 expression levels with RT-PCR and caspase-3 activity were analyzed using a kit. For detection of apoptotic activity, an in situ TUNEL assay was performed. Results: When 98.3µM ?-VNF, 52.5µM VNC and the 11.25+15.8µM VNC+?-VNF combination were compared with the control group, ??m changes at the 6th hour were found to be 19.5%, 5.5%, 24.6%, and 3.5% , respectively. These finding show that the combination group (24.6%) resulted in early apoptosis of the cell at the 6th hour. Bax mRNA expression increased at the 24th hour in the VNC+?-VNF group compared to control group (160%), and caspase-3 activation increased in the 1.25+15.8 µM[VNC+?-VNF] group compared to the control group at the 48th hour. The detection of DNA fragments in HepG2 cells within 24 hours suggests direct apoptosis. Conclusion: These findings demonstrate that the doses administered to the VNC+?-VNF combination group were lower than those administered to groups using one agent alone (e.g. VNC), the results of which reduce the possibility of administering toxic doses.

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