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

Abstract

Objective: This study was designed to investigate the effects of ε-viniferin (ε-VNF) on the mitochondrial pathway of apoptosis and on late apoptosis in HepG2 cell lines. To observe these effects, ε-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 (ΔΨm) were evaluated using cationic dye JC-1, while Bax, Bcl-2 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 ε-VNF, 52.5µM VNC and the 11.25+15.8µM VNC+ε-VNF combination were compared with the control group, ΔΨm 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+ε-VNF group compared to control group (160%), and caspase-3 activation increased in the 1.25+15.8 µM[VNC+ε-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+ε-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.

Keywords

• viniferin,vincristine,HepG2 cell

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