The Mechanisms of Chansu in Inducing Efficient Apoptosis in Colon Cancer Cells
Figure 2
CBF-induced apoptosis is both caspase-3 dependent and independent in HCT116 cells. (a) Expression of active caspase-3 detected by confocal microscopy. HCT116 cells were treated with 0 or 1 µM of CBF for 24 hours and analysed by immunofluorescence. The expression of active caspase-3 (green fluorescence) was only observed in treated HCT116. Cell nuclei were stained with DAPI (blue fluorescence). Scale bars equal 10 μm. (b) Detection of caspase-3/7 activity via a time course. A significant increase of caspase-3/7 activity was found after 24-hour-CBF exposure (, * compared to the control group). (c) RT-PCR of caspase-3 and AIF. While caspase-3 transcript levels decreased at initial 12 hours but increased at 24 hours, the mRNA level of AIF decreased by time. Results are means with standard errors from 3 replicates. (d) Protein expression of active caspase-3 and cleaved AIF after CBF exposure. After 24-hour treatment, both effective caspase-3 (17 kDa) and dissociated AIF (57 kDa) were produced. (e) Enhancement of cancer cell survival by NAC. The mixture of different concentrations of CBF and NAC partially inhibited cell death (, *).
