论文部分内容阅读
Aim:To elucidate the mechanism responsible for the antiproliferative effects of anovel homospermidine conjugate,anthracenylmethyl homospermidine(ANTMHspd),in the human hepatoma BEL-7402 cell line.Methods:The viabilityof the cells was assessed by MTT assay and the trypan blue dye exclusion method.Morphological changes were observed by fluorescence microscopy with Hoechst33258 staining.Cell cycle distribution,apoptosis,and mitochondrial membranepotential were measured by flow cytometry.Protein expression was detected byWestern blot analysis.Results:ANTMHspd strongly decreased BEL-7402 cellproliferation in a dose-and time-dependent manner.Hoechst 33258 staining andthe flow cytometry assay showed that ANTMHspd induced cell apoptosis andcell cycle perturbation.Furthermore,ANTMHspd could induce mitochondrialmembrane potential loss and cytochrome c release and enhance cleaved caspase-3,cleaved caspase-9,and Bax protein expression without caspase-8 activation.ANTMHspd could also decrease the expression of Bcl-2 and cytochrome c inmitochondria.In addition,the specific inhibitors of caspase-9 and caspase-3almost abolished the ANTMHspd-induced caspase-9 and caspase-3 activation,respectively.Conclusion:ANTMHspd could induce BEL-7402 cell apoptosis viathe mitochondrial/caspase-dependent pathway and the Bcl-2 family was involvedin the control of apoptosis.
Aim: To elucidate the mechanism responsible for the antiproliferative effects of anovel homospermidine conjugate, anthracenylmethyl homospermidine (ANTMHspd), in the human hepatoma BEL-7402 cell line. Methods: The viability of the cells was assessed by MTT assay and the trypan blue dye exclusion method . Morphological changes were observed by fluorescence microscopy with Hoechst 33258 staining. Cell cycle distribution, apoptosis, and mitochondrial membrane potential were measured by flow cytometry. Protein expression was detected by Western blot analysis. Results: ANTMHspd strongly decreased BEL-7402 cellproliferation in a dose-and time -dependent manner. Hoechst 33258 staining and the flow cytometry assay showed that ANTMHspd induced cell apoptosis and cell cycle perturbation. Frtherrtherm, ANTMHspd could induce mitochondrialmembrane potential loss and cytochrome c release and enhance cleaved caspase-3, cleaved caspase-9, and Bax protein expression without caspase-8 activation.ANTMHspd could also decrease the e xpression of Bcl-2 and cytochrome c in mitochondria. In addition, the specific inhibitors of caspase-9 and caspase-3 are mostly abolished the ANTMHspd-induced caspase-9 and caspase-3 activation, respectively. Confluence: ANTMHspd could induce BEL- 7402 cell apoptosis viathe mitochondrial / caspase-dependent pathway and the Bcl-2 family was involvedin the control of apoptosis.