RESUMO
Combined treatment with the medicinal mushroom Ganoderma lucidum and the herb Duchesnea chrysantha extracts (GDE) causes a synergistic induction of mitochondrial damage and apoptosis in HL-60 cells. GDE treatment is selectively toxic to HL-60 leukemia cells whereas no cytotoxic effect is observed in normal peripheral blood mononuclear cells. GDE-induced apoptosis is associated with Bcl-2 down-regulation, Bax translocation, mitochondrial cytochrome c release and caspase-3 activation, suggesting that apoptosis by this combination occurs through the mitochondria-dependent pathway. The present findings suggest that this combination merits further investigation as a potential therapeutic agent for the treatment of cancer.
Assuntos
Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Basidiomycota/química , Mitocôndrias/efeitos dos fármacos , Rosaceae/química , Clorometilcetonas de Aminoácidos/farmacologia , Antineoplásicos/química , Western Blotting , Caspase 3/metabolismo , Inibidores de Caspase , Sobrevivência Celular/efeitos dos fármacos , Citocromos c/metabolismo , Relação Dose-Resposta a Droga , Ativação Enzimática/efeitos dos fármacos , Citometria de Fluxo , Inibidores do Crescimento/química , Inibidores do Crescimento/farmacologia , Células HL-60 , Humanos , Leucemia/metabolismo , Leucemia/patologia , Mitocôndrias/metabolismo , Membranas Mitocondriais/efeitos dos fármacos , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Transporte Proteico/efeitos dos fármacos , Fatores de Tempo , Proteína X Associada a bcl-2/metabolismoRESUMO
BACKGROUND: We previously showed that an Epstein-Barr virus (EBV)-based plasmid, pEBVGFP, exerts prolonged gene expression in porcine neonatal pancreatic cell clusters (NPCCs). In this study, the mechanism underlying this was investigated. METHODS: GFP expression was analyzed in porcine cells transfected with pEBVGFP by FACS analysis and confocal microscopy. The possible integration of pEBVGFP into the chromosomal DNA was analyzed by Southern blot. Self-replication of the EBV-based plasmid in porcine cells was investigated by PCR. The NPCCs were immunostained to characterize cells transfected with pEBVGFP. RESULTS: The EBV based plasmid provided prolonged GFP expression in porcine cells and duct cells were the main cells transfected among NPCCs. Southern blot showed that the transfected pEBVGFP stayed for a long time as an episome rather than integrating into the chromosomal DNA. pEBVGFP isolated from the transfected porcine cells had methylated CpG suggesting that they self-replicated in those cells. CONCLUSIONS: The EBV-based plasmid may be useful for genetically manipulating porcine cells to enhance their value as xenotransplantation sources.