title: A novel homospermidine conjugate inhibits growth and induces apoptosis in human hepatoma cells.
abstract: OBJECTIVE
  To elucidate the mechanism responsible for the antiproliferative effects of a novel 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 cell proliferation 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. Furthermore, ANTMHspd could induce mitochondrial membrane 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 in mitochondria. In addition, the specific inhibitors of caspase-9 and caspase-3 almost abolished the ANTMHspd-induced caspase-9 and caspase-3 activation, respectively.
  
  
  CONCLUSIONS
  ANTMHspd could induce BEL-7402 cell apoptosis via the mitochondrial/caspase-dependent pathway and the Bcl-2 family was involved in the control of apoptosis.
authors.affiliation: Department of Pharmacology, Shenyang Pharmaceutical University, Shenyang 110016, China.
authors.forename: Song-qiang
authors.initials: SQ
authors.lastname: Xie
authors.forename: Ying-liang
authors.initials: YL
authors.lastname: Wu
authors.forename: Peng-fei
authors.initials: PF
authors.lastname: Cheng
authors.forename: Min-wei
authors.initials: MW
authors.lastname: Wang
authors.forename: Guang-chao
authors.initials: GC
authors.lastname: Liu
authors.forename: Yuan-fang
authors.initials: YF
authors.lastname: Ma
authors.forename: Jin
authors.initials: J
authors.lastname: Zhao
authors.forename: Chao-jie
authors.initials: CJ
authors.lastname: Wang
mesh_terms: D000873:Anthracenes / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy* / Q000473:pathology*; D020169:Caspases / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D045304:Cytochromes c / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D006801:Humans; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D013095:Spermidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D013997:Time Factors; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: D000873:Anthracenes; D019253:Proto-Oncogene Proteins c-bcl-2; C529696:anthracenylmethyl homospermidine; D051028:bcl-2-Associated X Protein; D045304:Cytochromes c; D020169:Caspases; D013095:Spermidine