Oleanolic acid and ursolic acid inhibit proliferation in transformed rat hepatic oval cells

Figure 2 H₂O₂ induced WB-F344 malignant transformation. A: The morphology of H₂O₂-treated WB-F344 cells. The cells were cultured in complete medium (control) or treated with 7 × 10^-7 mol/L H₂O₂ 12 h per day for 21 d. Cell morphology was observed under microscope (× 400). An increasing nucleus to cytoplasm ratio was observed (green arrow), as were many mitotic cells (yellow arrow), prokaryotes (blue arrow) and even tumor giant cells (red arrow); B: The tumorigenicity of transformed WB-F344 cells as determined using the methylcellulose culture assay. Cell colony formation was observed under microscope (× 400); C: H₂O₂ increased WB-F344 cell aneuploidy. The cells were cultured in complete medium (control) or treated with 7 × 10^-7 mol/L H₂O₂ 12 h per day for 21 d and harvested for flow cytometry. DNA was stained with propidium iodide (10 μg/mL), and a BD FACS Calibur was used to analyze cellular DNA content. The population of > 4N cells represent aneuploidy cells. Histograms indicating the aneuploidy cells among the control cells and the H₂O₂-treated cells. The data represent the mean ± SD derived from three independent experiments.