Apoptosis Induction in Cancer Cells

Aqueous, 50% and 100% ethanol Tritticum spelta and Salix mucronata plant extracts were tested for their pro-apoptotic activity. The annexin V apoptosis detection kit for flow cytometry (Sigma-Aldrich, MO, USA) was used. The annexin V assay was carried out in conjunction with propidium iodide (PI) staining. HepG2 and Caco-2 cancer cell lines were cultured for 24 h in a 25 cm² culture flask (1 × 10⁶ cells/well) with 66.6 μg/mL (the lowest IC₅₀) of the investigated extracts. After 72 h, cells were harvested by trypsinization and centrifuged at 1000 rpm for 5 min and then re-suspended in 1 × binding buffer prior to staining with 5 μL of annexin V and 10 μL of propidium iodide solution for 15 min at room temperature. The apoptosis-dependent anti-proliferative effect was determined by quantification of annexin-stained apoptotic cells using the FITC signal detector (FL1) against the phycoerythrin emission signal detector (FL2)^{20 (link)}.