Mechanism Of Apoptosis Induced By Ursolic Acid On Human Hepatic Carcinoma BEL-7404 Cells

Objective:To investigate the anti-tumor effect and apoptosis-inducing mechanism of UA on human hepatic carcinoma BEL-7404 cells. Methods:Based on the culture of human hepatic carcinoma BEL-7404 cells in vitro. Control group, UA group and DDP group were included. The effect of ursolic acid on growth of human hepatic carcinoma BEL-7404 cells was evaluated by MTT. To observe occurrence of apoptosis in the early, intermediate and terminal stage, the morphology, DNA fragmentation and phos- phatidylserine(PS) residues were detected by transmission electron microscopy, agarosegel electrophoresis and flow cytometry, respectively. Procaspase-3, caspase-9 and survivin proteins expression obtained by Western Blot analysis. Results:①MTT showed UA had a significant anti-proliferation effect on human hepatic carcinoma BEL-7404 cells,UA50μmol/L and DDP10μg/ml had the best anti-proliferation effect after 48h as the experiment condition.②Transmission electron microscopy showed early apoptosis cells had no villus, vacuole inside, the terminal stage apoptosis cells showed nucleus condensation and apoptotic bodies.③Hoechst33258 showed chromatin condensation with fluorescence after incubation by UA.④Result of flow cyto-metry showed apoptosis rate was increased after treatment with UA and DDP for 48h (F=3698.99, P<0.01).⑤Western Blot analysis indicated that expression level of procaspase-3 and survivin was descend, but caspase-9 increased. Conclusions:UA has the effect of anti-proliferation and inhibits apoptosis on BEL-7404 cells, the mechanism is that human hepatic carcinoma BEL-7404 cells were treated by UA can increase expression of caspase-9, but elevate the expression of procaspase-3, which can disjoint DNA, matrix proteins and framework proteins, at the same time, inhibit survivin activity, low down sensitivity to the factors of apoptotis, which intensify cell apoptosis.