Apoptosis Kinetics in Colon and Rectum Cancer

The levels of cell surface phosphatidylserine were determined to monitor the type of cell death and the kinetics of apoptosis [17 (link)]. The CRC cell lines SW1116 and SW837 were seeded (2.5 × 10⁵ cells/well) into 24-well plates and incubated in a non-CO2 incubator at 37˚C for 18 h. Colon cancer (SW1116) cells were simultaneously treated with Sora (5 µM) and Cur or Que (200 and 400 µM) for 72 h, and rectum cancer (SW837) cells were treated with Sora (5 µM) and Cur or Kmf (200 and 400 µM). The cells were subsequently washed twice with HBSS and harvested using trypsin. Finally, the cells were double-stained using the Annexin V-FITC-Flous staining kit, according to the manufacturer’s instructions (Roche Diagnostic GmbH). Annexin V-Flous labelling solution containing Annexin V-FITC and PI (100 µl) was added to both the treated and control cell groups and incubated at 15–20˚C for 15 min. The cells (1 × 10⁶ cells/ml) were then resuspended in binding buffer, and fluorescence was monitored using flow cytometry (FC500; Beckman Coulter, Inc.).