骨肉瘤_____Survivin；

Survivin基因干扰RNA对骨肉瘤MG63细胞系增殖和凋亡的影响

http://www.100md.com 《中国矫形外科杂志》 2006年第5期

骨肉瘤,,Survivin；,siRNA；,短发夹状RNA；,骨肉瘤；,细胞增殖；,细胞凋亡,1材料与方法,2结果,3讨论

     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 21neoSurvivin，转染骨肉瘤细胞系MG63，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，RTPCR、Westonblot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MG63细胞Survivin基因mRNA水平和蛋白表达显著下降，其抑制率分别为8508%和8114%；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6341%；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2454%。[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

    Effect of a short hairpin RNA targeting Survivin on the proliferation and apoptosis in osteosarcoma cell line MG63∥LI Kun,YANG Shuhua,LIU Hongyun,et alOrthopaedic Department,Union Hospital Affiliated to Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430022

    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 MG63 in vitro[Method]A plasmid of a short hairpin RNA targeting Survivin was constructed,and it was transfered into MG63 cell line by dosper liposomal methodMorphological change of tumor cells was observed under invert microscopeThe expression of Survivin mRNA and protein were examined by RTPCR and Westonblot method respectivelyThe inhibition of the cell proliferation was estimated by MTT method and the colony forming testThe apoptotic cells were stained by acridine orange[Result]The number of drift cells was increased after transfection,Survivin shRNA could significantly reduce the expressions of Survivin mRNA and proteinThe expression of Survivin mRNA and protein after transfection was respectively inhibited by 8508% and 8114%,Survivin shRNA also inhibit the growth of the cell by MTT method and the inhibition rate reached 6341% at 48 h,the apoptosis rate of the cells in the transfected group was 2454%[Conclusion]Survivin shRNA could significantly reduce the expressions of Survivin mRNA and protein and inhibit the proliferation of the MG63 cell,and increase the apoptosis of the cells ......

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