Title: Investigation of apoptosis inducing effect of ponicidin on leukemia NB4 cells in vitro
Abstract: OBJECTIVE:To investigate the anti-proliferation and apoptosis inducing effects of ponicidin (PON) on leukemic NB4 cells and its mechanisms of action.METHODS:NB4 cells in culture medium in vitro were given with different concentrations of PON (0-50 μmol/L) for 0,24,48 and 72 h.The inhibitory rate of the cells was measured by MTT assay and cell apoptosis was examined by flow cytometry (FCM).Western blotting was used to detect Caspase-3 and poly(ADP-ribose) polymerase (PARP) expression as well as apoptotic modulators such as Bax,Bcl-2,Bak,Bad and Survivin.RESULTS:PON (over 20 μmol/L) inhibited the growth of NB4 cells in both time-and dose-dependent manners.The cell apoptosis was demonstrated obviously by FCM and marked morphological changes of cell apoptosis such as the condensation of chromatin was clearly observed by Hoechst staining after treatment for 48 h.Western blotting showed cleavage of the Caspase-3 zymogen protein,with the appearance of its 17×103 subunit,and a cleaved 89×103 fragment of PARP was also found.Furthermore,Western blotting showed that anti-apoptotic protein Survivin decreased dramatically while pro-apoptotic protein Bax increased significantly after the cells were treated by PON for 48 h.CONCLUSIONS:PON exhibits in vitro anti-leukemia effect by induction of apoptosis in NB4 cells,and downregulation of Survivin as well as upregulation of Bax expression may be one of its significant apoptosis inducing mechanism.
Publication Year: 2010
Publication Date: 2010-01-01
Language: en
Type: article
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