Figure 3. Apoptotic cell death and expression of apoptosis-related proteins of colon cancer cells by inflexinol. A, colon cancer cells were treated with several concentrations of inflexinol for 24 h; cell morphological changes were observed under a microscope (top); and apoptotic cells were examined with a fluorescence microscope after TUNEL staining (bottom). Total number of cells in a given
Figure 3. Apoptotic cell death and expression of apoptosis-related proteins of colon cancer cells by inflexinol. A, colon cancer cells were treated with several concentration of inflexinol for 24 h; cell morphologic changes were observed under a microscope (top); and apoptotic cells were examined with a fluorescence microscope after TUNEL staining (bottom). Total number of cells in a given area was determined by DAPI nuclear staining (fluorescent microscope; middle). The apoptotic index was determined as the number of DAPI-stained TUNEL-positive cells counted. Columns, mean of three experiments each done in triplicate; bars, SD. *, P < 0.05, versus the untreated group. B, the cells were treated with different concentrations (10–40 μmol/L) of inflexinol at 37°C for 24 h. Equal amounts of total proteins (50 μg/lane) were subjected to 12% SDS-PAGE. Expression of Bax, cleaved caspase-3, cleaved caspase-9, cleaved poly(ADP-ribose) polymerase (PARP), cyclin D1 (CyD1), Bcl-2, XIAP, cIAP1/2, and β-actin were detected by Western blotting using specific antibodies. β-Actin protein was used as an internal control. Each blot is representative of three independent experimental results. Bar, 100 μm.