pSilencer-VEGF-siRNA诱导骨肉瘤细胞系MG63凋亡并上调caspase-3表达

Upregulation of caspase-3 expression and apoptosis of MG63 cell line induced by pSilencer-VEGF-siRNA transfection

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—72h，流式细胞仪AnnexinV-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G。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pSilencer-VEGF-siRNA可诱导MG63细胞早期凋亡；并可上调caspase-3的表达。

Objective To research effects on caspase-3 expression of osteosarcoma cell line MG63 induced by pSileneer-VEGF-siRNA transfection. Methods The cell line MG63 was cultured and human-specific pSilencer-VEGF- siRNA was constructed. The experiment was divided into pSileneer-VEGF-siRNA transfeetion 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.