Induction of apoptosis and cell cycle arrest in human HCC MHCC97H cells with Chrysanthemum indicum extract

Figure 1 Effects of Chrysanthemum indicum extract (CIE) on cell viability of normal cells. A: Rat hepatocytes; B: Human umbilical vein endothelial cell line ECV304. Various concentrations of CIE were added, and the cells were incubated for 24 h. Cell viability was measured by 0.2% trypan blue exclusion (A) and MTT assay (B) respectively. Results presented are representative of 3 independent experiments.

Figure 2 Dose-response curves for CIE in MHCC97H cells, following 24 h, 48 h and 72 h incubation, as assessed by MTT assay. CIE produced a concentration- and time-dependent decrease in cellular proliferation. Results presented are representative of 3 independent experiments.

Figure 3 Effect of CIE on the induction of MHCC97H cell apoptosis. A: The apoptosis of MHCC97H cells induced by CIE were determined by flow cytometry at 24 h; B: CIE induced a concentration-dependent increase in early and late cellular apoptosis. Results presented are representative of 3 independent experiments. ^aP < 0.05, ^bP < 0.01 vs control group.

Figure 4 Morphological changes in MHCC97H cells with CIE. Cells were observed by phase contrast microscopy in controls and after treatment with 400, 800, 1200 μg/mL CIE (× 200).

Figure 5 Effect of CIE on MHCC97H cellular mitochondrial membrane potential (MMPΔψ_m). A: The MMPΔψ_m of MHCC97H cells were determined by flow cytometry at 24 h after CIE; B: Results presented are representative of 3 independent experiments. ^aP < 0.05, ^bP < 0.01 vs control group.

Figure 6 Effect of CIE on apoptosis-related protein expression. A: The expressions of cytochrome C, caspase-9 and caspase-3 were assessed by Western blotting; B: Results presented are representative of 3 independent experiments. ^aP < 0.05, ^bP < 0.01 vs control group.

Figure 7 Effect of CIE on MHCC97H cell cycle. A: The cell cycle of MHCC97H cells in the presence of CIE was determined by flow cytometry at 48 h; B and C: Results presented are representative of 3 independent experiments. ^aP < 0.05, ^bP < 0.01 vs control group.

Figure 8 Effect of CIE on cell cycle-related protein expression. A: The expression of CDK4 and P21 was assessed by Western blotting; B: Results presented are representative of 3 independent experiments. ^bP < 0.01 vs control group.