【摘要】 目的设计并合成抗人骨肉瘤Periostin基因的siRNA分子,观察基因沉默效应。方法根据GenBank上获取的Periostin mRNA序列合成4条siRNA分子,用脂质体介导转染体外培养的肉瘤细胞MG-63细胞,培养48h,采用ELISA评估、RT-PCR检测和Western blot检测分析基因沉默效果。结果不同实验组之间的蛋白表达水平的差异有统计学意义(F=86.52,P<0.01),与对照组相比,4条Periostin siRNA均下调Periostin蛋白表达的水平,其中以Periostin siRNA3效果最好,抑制率分别为58.06%、61.78,75.33%和51.67%。结论靶向Periostin siRNA均有效抑制Periostin蛋白的表达水平。更多还原

【Abstract】 Objective Nine periostin siRNAs were designed and the silencing efficiency was evaluated.Methods Four periostin siRNA sequences were designed and transfected into human osteosarcoma MG-63 cells by lipofectamine RNAiMAX reagent,the transcription level of periostin was analyzed by ELISA,qRT-PCR and Western blotting.Results The three periostin specific double stranded siRNA inhibited the expression of periostin.The inhibiting rates of periostin siRNA1/siRNA2/siRNA3/siRNA4 were 58.06%,61.78%,75.33%,and 51.67%,respectively.Conclusion Periostin siRNAs was successfully designed and synthesized,which can specifically and effectively downregulate expression of periostin gene.更多还原