An Experiment Study On Inhibition Of Eleo-Zedoray Parenteral To RL-952,HepG₂ In Vitro

Objective To investigate the effects of Eleo-Zedoray parenteral solution on cell cycle, proliferation and metastasis of RL-952 and HepG2 cells in vitro and the opportune mechanism.Methods RL-952 and HepG2 cells were treated with different concentration of Eleo-Zedoray parenteral solution. By using the MTT assay to display the paste rate and the sensitivityof RL-952 and HepG2 cells to the drug, the inhibition of Eleo-Zedoray parenteral solution to them in vitro was observed. Morphologic changes were studied under Light Microscope (LM) and transmission Electron Microscopy (EM) . We using Flow cytometry (FC) to assay cell-DNA distribution and apoptosis. By immunocytochemical method, the expressions of CD1, CE, P16, P53, Fas, Caspase-3 and E-cad were observed.Result The inhibition rate of Eleo-Zedoray parenteral solution to RL-952 and HepG2 indicated dose-dependent and time-dependent anti-proliferative effect. Treated with 1/2 IC50 of Eleo-Zedoray parenteral solution on RL-952 and HepG2 cells for 24 hours, 48 hours, 72 hours, cell cycle progression was delayed at G0/G1 phase. Apoptosis was confirmed by the appearance of apoptosis peak on FCM and characteristic ultrastructural changes on EM in RL-952 cells. After treatment the expression of cellcycle negative regulation factor, P16 ,was increased . And the paste rate experiment showed the paste rate decreased obviously after treated with 1/2 IC50 of Eleo-Zedoray parenteral solution on RL-92 and HepG2 cells for 72 hours, .Conclusion Eleo-Zedoray parenteral solution can inhibit the proliferation of RL-952 and HepG2 cells and increase proportion of the G0/G1 phase cells. The effect is related to the up-regulation of P16. It also induces the apoptosis by increasing caspase-3 activity, but not the Fas receptor in RL-952 cells. Eleo-Zedoray parenteral solution can decrease the paste rate of RL-952 and HepG2 cells.