To further confirm that C. orbiculata induces apoptosis in HCT116 colon cancer cells, we used Annexin-V-FITC and PI using flow cytometry to quantify the percentage of viable, apoptotic, late apoptotic and necrotic cells in HCT116 cells treated with varying concentrations of C. orbiculata. This evidence concerns the gene ANXA5 and malignant colon neoplasm.