Mechanisms of apoptosis induced by PBA-D1 in Hela cells

Aim To study whether the derivation of pseudolaric acid B(PBA-D1) can induce apoptosis on human galactophore cancer Hela cells line in vitro and the possible mechanisms of apoptosis. Methods MTT,SRB assay,cell growth curve,colony formation of cancer cells, morphological study, DNA gel electrophoresis, flow cytometry,Western blot.Results Potent cytotoxic effect on human galactophore cancer Hela cells line can be observed by MTT,SRB assay and cell growth curve,colony formation of cancer cells. After treatment of Hela cells with PBA-D1, apparent morphological characteristic of apoptosis was detected under the fluorescent microscopes with the staining of Hoechst 33342, both the number of apoptosis cells and the cell circle were measured by flow cytometry, a typical “Sub-G_1 peak” was also checked and PBA-D1 blocked the tumor cells at G_2+M phase and arrested the following cell circle. Western blots showed PBA-D1 promoted p53 protein expression, at the same time it also inhibited bcl-2 expression in a dose-dependent manner.Conclusion PBA-D1 induces apoptosis by promoting p53 and inhibiting bcl-2 expression in a dose-dependent manner.