靶向人骨肉瘤Periostin基因siRNA的构建及其沉默效应  

Construction of Small Interfering RNA Targeting Human Osteosarcoma Periostin Receptor and Gene Silencing Efficiency

作　　者：胡飞[1] 尚希福[1] 王伟[2] 周航成[3] 

机构地区：[1]安徽省立医院骨科,合肥230001 [2]安徽省立医院肿瘤科,合肥230001 [3]安徽省立医院病理科,合肥230001

出　　处：《临床输血与检验》2015年第2期122-125,共4页Journal of Clinical Transfusion and Laboratory Medicine

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。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.

关 键 词：小干扰RNA 骨膜蛋白 骨肉瘤 构建 

分 类 号：R392.11[医药卫生—免疫学] R738.1[医药卫生—基础医学]