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Survivin基因干扰RNA对骨肉瘤MG
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【防疫医学论文】作者:李鲲,杨述华,刘红云,吴强,傅德皓【关键词】 骨肉瘤摘 要:[目的]观察Survivin基因siRNA表达载体对骨肉瘤细胞增殖和凋亡的 影响 。[ 方法 ]体外构建Survivin shRNA表达载体pSilence 21neoSurvivin,转染骨肉瘤细胞系MG63,筛选稳定表达的克隆,倒置显微镜观察各组细胞形态学变化,RTPCR、Westonblot方法检测转染后Survivin mRNA及蛋白的表达,噻唑蓝(MTT)法、克隆形成实验检测细胞的增殖情况,吖啶橙荧光染色法观察细胞凋亡情况。[结果]转染后MG63细胞Survivin基因mRNA水平和蛋白表达显著下降,其抑制率分别为8508%和8114%;细胞增殖受到显著抑制,培养48 h后与空白组比较,抑制率达6341%;吖啶橙染色显示转染组细胞出现明显核碎裂等凋亡改变,转染组凋亡率为2454%。[结论]靶向Survivin基因的siRNA表达载体可以显著抑制骨肉瘤细胞增殖,促进细胞凋亡。 关键词:Survivin; siRNA; 短发夹状RNA; 骨肉瘤; 细胞增殖; 细胞凋亡 Abstract:[Objective]To construct an expression plasmid of a short hairpin RNA targeting Survivin,and investigate its effect on the expression of Survivin,the proliferation and the apoptosis of human osteosarcoma cell line MG63 in vitro[Method]A plasmid of a short hairpin RNA targeting Survivin was constructed,and it was transfered into MG63 cell line by dosper liposomal methodMorphological change o
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