【摘要】 目的研究褪黑素对人骨肉瘤细胞survivin、Caspase-3表达的调控,进一步探讨褪黑素的抗肿瘤机制。方法体外培养人骨肉瘤细胞MG63、U2,Hoechst33258染色观察两种骨肉瘤细胞凋亡的改变,实时荧光定量PCR及免疫印迹法检测两种骨肉瘤细胞survivin、Caspase-3表达。结果 Hoechst33258染色显示褪黑素作用后促进骨肉瘤细胞MG63、U2凋亡,实时荧光定量PCR及免疫印迹分析结果表明,经褪黑素作用后的骨肉瘤MG63、U2细胞中survivin在mRNA水平表达的无明显差别,但其蛋白质水平较用药前明显降低;Caspase-3在mRNA及蛋白质水平上均较用药前明显增高。结论褪黑素诱导人骨肉瘤细胞MG63、U2发生凋亡,可能是通过抑制survivin基因在蛋白水平的表达,同时上调Caspase-3基因的表达,进而促进细胞凋亡,发挥其抗肿瘤作用。更多还原

【Abstract】 Objective To study the regulation of the melatonin on expression of survivin and Caspase-3 in human osteosarcoma cells and to further explore the anti-tumor mechanism of melatonin. Methods Human osteosarcoma cells MG63 and U2 were cultured in vitro. Hoechst33258 staining was used to observe the apoptosis of two osteosarcoma cells. The expressions of survivin and Caspase-3 genes of the two osteosarcoma cells were detected by real-time fluorescence quantitative PCR and western blot. Results Hoechst33258 staining showed that the melatonin promoted apoptosis of MG63 and U2 osteosarcoma cells. The results of real-time fluorescence quantitative PCR and western blot showed that the expressions of survivin gene in MG63 and U2 cells of osteosarcoma after melatonin treatment were not significantly different in mRNA level, but significantly lower in protein level than those in cells before treatment(P<0.05). The expressions of Caspase-3 gene were significantly higher in both mRNA and protein levels than those in cells before treatment(P<0.05). Conclusion Melatonin induces apoptosis of osteosarcoma cells MG63 and U2 possibly by inhibiting survivin gene expression in protein level and up-regulating Caspase-3 gene expression, which promotes apoptosis and exert its anti-tumor effects.更多还原