| Objective：To investigate the effects of TSCO on the proliferation and apoptosis inthe leukemia cell line(K562) and the possible mechanisms were tried to explored.Method：(1) The optimal parameters of the total saponin extraction from the cornus byethand refluxing method, HPD-100macroporous resin shows better performance forpurifying the cornus total saponin.(2) MTT assay was used to examine the growth inhibition of differentconcentration for different time.(3) The apoptosis of K562cells induced by the drug were investigated by meansof DNA transmission electron microscope and cell nucleus’morphology.(4) The expressions of Caspase-3, Bax and Bcl-2in K562cells acted by TSCOwere analyzed by spectrophotom etry and Western bloting.Results:(1)With the growth of concentration of TSCO, it began to inhibit K562theproliferation of cells.The higher concentration and the more time, the more cells wereinhibited.(2) The characteristics of apoptosis in K562cells induced by TSCO wereobserved: reduction of nucleus, chromatin condensation and nuclear membranerupture; And with TSCO at different dosage for72h, apoptotic cells are increasesignificantly(P<0.01) by means of cellnucleus’ morphology.(3) A typical ladder pattern was identified in DNA electrophoresis after TSCOacted on K562for72h.(4) After TSCO acted on K562for72h, the expressions of Caspase-3in K562cell were inhibited., compared with the control group, the difference issignicant(P<0.01).(5) The expressions of Bax were inhibited, and Bcl-2expression decreasedobviously after the K562cells were treated with TSCO for72h.Conclusion:(1)TSCO can inhibit cells proliferate significantly and the inhibition take on a dose-and time-dependent manner.(2)TSCO can induce K562cells apoptosis and the mechanism of apoptosis maybe related to up-regulation of Caspase-3, Bax and down-regulation of Bcl-2expression. |
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