YB-13,a novel synthetic microtubule inhibitor,induces apoptosis of HeLa cells and its mechanism

Aim To study whether the derivative of indol-3-glyoxylic acid amide induced apoptosis on human galactophore cancer HeLa cells line(HeLa cells)in vitro and the feasible mechanisms of apoptosis.Methods MTT assay,cell growth curve,colony formation of cancer cells,morphological study,DNA gel electrophoresis,flow cytometry,RT-PCR,and fluorescence microscopy were carried out.Results Potent cytotoxic effect on HeLa cells could be observed by MTT assay and cell growth curve,colony formation of cancer cells.After treatment of HeLa cells with YB-13,the apparent morphological characteristic of apoptosis was detected under the fluorescent microscopes by the staining of Hoechst 33342.Both the number of apoptosis cells and the cell circle were measured by flow cytometry.A typical Sub-G1 peak was also checked and YB-13 blocked the tumor cells at G2+M phase and arrested followed cell circle.RT-PCR showed that YB-13 promoted bax mRNA expression,at the same time it also inhibited bcl-2 expression in a dose-dependent manner.YB-13 can interfere with microtubule polymerization and disrupting cytoskeleton.Conclusions YB-13 shows obvious anticancer activity by inducing apoptosis and causing cell cycle arrest in a dose-dependent manner and can interfere with microtubule polymerization and disrupting cytoskeleton.