Fucoidan induces apoptosis of HT-29 and HCT116 cells. (A) HT-29 cells were plated on cell culture coverslips at 50,000 cells/cover and treated for 72 h with fucoidan, as shown in Figure 1. The cells were stained with Hoechst 33258 and the microphotograph images are representative of three independent experiments. Magnification, × 200. (B) HT-29 cells were treated with or without 20 μg/mL of fucoidan for the indicated periods as described in Figure 1. HT-29 (C) and HCT116 (D) cells were treated with various concentrations of fucoidan for 72 h. (B, C, D) The cells were trypsinzed, stained with 7-amino-actinomycin D and annexin V, and then analyzed via flow cytometry. The numbers of living cells and early apoptotic cells are expressed as percentages of the total cell number. Results are expressed as the means ± SEM (n = 6). HT-29 (E), HCT116 (F), and FHC (G) cells were plated in 100 mm dishes and treated with various concentrations of fucoidan for 60 h, as described in Figure 1. The cell lysates were analyzed via Western blotting with the indicated antibodies. A photograph of chemiluminescent detection of the blots, which were representative of three independent experiments, are provided. The relative abundance of each band to its own β-actin was quantified, and the control levels were set to 1. The adjusted mean ± SEM (n = 3) of each band is shown above each blot. (B) *Significantly different from 0 μg/mL of fucoidan, P < 0.05. (C, D, E, F) Means without a common letter differ, P < 0.05.
Kim et al. BMC Gastroenterology 2010 10:96 doi:10.1186/1471-230X-10-96