【摘要】 目的研究pSilencer-VEGF-siRNA转染骨肉瘤细胞系MG63后诱导其凋亡以及对caspase-3表达能力的影响。方法体外常规培养骨肉瘤细胞系MG63并构建人类特异性的pSilencer-VEGF-siRNA;分为pSilencer-VEGF-siRNA转染组和空载体组。转染后48~72 h,流式细胞仪Annexin V-FITC/PI染色检测细胞凋亡、分析细胞周期;Westernblot法测定caspase-3表达。结果MG63转染pSilencer-VEGF-siRNA表达质粒后,早期出现细胞凋亡及明显的G1期阻滞以及G1期细胞数增加;pSilencer-VEGF转染组caspase-3的表达较空载体组和对照组明显上调。结论pSilenc-er-VEGF-siRNA可诱导MG63细胞早期凋亡;并可上调caspase-3的表达。更多还原

【Abstract】 Objective To research effects on caspase-3 expression of osteosarcoma cell line MG63 induced by pSilencer-VEGF-siRNA transfection.Methods The cell line MG63 was cultured and human-specific pSilencer-VEGF-siRNA was constructed.The experiment was divided into pSilencerVEGF-siRNA transfection and vector group.Apoptosis and cell cycle of MG63 was detected and analysed with conjugated annexin-V(Annexin-V-FITC) antibody and propidium iodide(PI) 48～72 hours with flow cytometry after transfection.The expression of caspase-3 was detected with Western blotting.Results The early apoptosis of MG63 was found after transfection and with a blockage of G1 phase and cell accumulation.Caspase-3 expression was up-regulated apparently in the transfection group as compared with vector group.Conclusion Early apoptosis of MG63 can be induced by pSilencer-VEGF-siRNA which up-regulates caspase-3 expression.更多还原