利用siRNA抑制人骨肉瘤细胞survivin的表达
Inhibition of the Expression of survivin in Osteosarcoma Cell by A Short Hairpin RNA
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摘要: 目的构建生存素(survivin)短发夹状RNA表达载体,检测其对骨肉瘤细胞survivin mRNA及蛋白表达的影响。方法体外构建survivin shRNA表达载体pSilence2.1-neo-survivin,转染骨肉瘤细胞系MG-63,RT-PCR、免疫组化方法检测转染前后survivin mRNA及蛋白的表达,MTT比色法检测细胞增殖活性。结果pSilence2.1-neo-survivin载体转染能显著抑制survivin mRNA、蛋白表达,转染后48h细胞增殖的抑制率为61.83%。结论pSilence2.1-neo-survivin可显著抑制MG-63细胞survivinmRNA、蛋白的表达及细胞的增殖活性。Abstract: Objective To construct a expression plasmid of a short hairpin RNA targeting survivin,and investigate its inhibitory effect on the expression of survivin in osteosarcoma cell line MG-63 in vitro. Methods Construct a expression plasmid of a short hairpin RNA targeting survivin,and transfect it into MG-63 cell by dosper liposomal.The expression of survivin mRNA and protein were examined by RT-PCR and immunohistochemical method,respectively.The inhibition of the cell proliferation was estimated by MTT method.Re...
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