摘要
The aim of this study is to investigate the anti-proliferative effects of Zebularine on caspase-3 and 9 genes by methylation and protein expression on SKBR3 cells. The SKBR3 cells were treated with Zebularine at different concentrations (0 - 140 μM) for 24 - 96 hours and the effects on the cell viability were shown. The effects on the cell migration and cell transformation were also demonstrated at the specified dose (IC50 = 40 μM). At the same time, the HRM method and western-blot analysis were used to understand the effects in the apoptotic mechanism. According to the obtained results, it was observed that Zebularine significantly decreased the cell proliferation, cell migration and the cell growth (p < 0.001). As a result of the methylation analyzes performed on SKBR-3 cells at the applied dose, it was observed that Zebularine caused a decrease in both caspase-3 and 9 methylation rates at the 72nd hour. However, both reductions in the methylation levels didn’t cause a significant change (p > 0.05). At the same time, it was detected by the western-blot analyses that there was a time dependent increase in caspase-3 genes while a decrease was detected in the caspase-9 gene in progress of time. The obtained results showed that the methylation changes occurring in the caspase genes weren’t related to the protein levels. According to these results, supportive results are obtained showing that the Zebularine can be used in chemotherapy and that this study is unique since it is the first study in the literature intending to investigate the effects of Zebularine on the SKBR3 cells.
The aim of this study is to investigate the anti-proliferative effects of Zebularine on caspase-3 and 9 genes by methylation and protein expression on SKBR3 cells. The SKBR3 cells were treated with Zebularine at different concentrations (0 - 140 μM) for 24 - 96 hours and the effects on the cell viability were shown. The effects on the cell migration and cell transformation were also demonstrated at the specified dose (IC50 = 40 μM). At the same time, the HRM method and western-blot analysis were used to understand the effects in the apoptotic mechanism. According to the obtained results, it was observed that Zebularine significantly decreased the cell proliferation, cell migration and the cell growth (p < 0.001). As a result of the methylation analyzes performed on SKBR-3 cells at the applied dose, it was observed that Zebularine caused a decrease in both caspase-3 and 9 methylation rates at the 72nd hour. However, both reductions in the methylation levels didn’t cause a significant change (p > 0.05). At the same time, it was detected by the western-blot analyses that there was a time dependent increase in caspase-3 genes while a decrease was detected in the caspase-9 gene in progress of time. The obtained results showed that the methylation changes occurring in the caspase genes weren’t related to the protein levels. According to these results, supportive results are obtained showing that the Zebularine can be used in chemotherapy and that this study is unique since it is the first study in the literature intending to investigate the effects of Zebularine on the SKBR3 cells.
