南方医科大学学报

南方医科大学学报 ›› 2015, Vol. 35 ›› Issue (03): 403-.

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高效能沉默BMP-7 基因siRNA序列的实验筛选

李彩霞，白鹏，赵娴，董苑，代晓明，李逸松，刘流   

  • 出版日期:2015-03-20 发布日期:2015-03-20

Screening of highly efficient small interfering RNAs against bone morphogenetic
protein-7

  • Online:2015-03-20 Published:2015-03-20

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摘要： 目的骨形态发生蛋白（bone morphogenetic protein-7, BMP-7）是一种促进骨和软骨再生的关键性生长因子，旨在筛选高
效能BMP-7siRNA序列，为后续探讨BMP-7的生物学作用机制的基础和临床应用研究提供实验研究技术手段。方法本实验
针对BMP-7基因，构建出相应4条siRNA序列，分别将其转染至大鼠脐静脉内皮细胞，转染72 h后用Wb和RT-PCR方法检测
BMP-7蛋白和BMP-7 mRNA表达水平，筛选出最高效的BMP-7siRNA序列。结果人工合成的4条BMP-7siRNA序列都能有
效抑制大鼠脐静脉内皮细胞的BMP-7表达，其中BMP-7-3siRNA序列抑制效果最为明显，抑制率大于90%，被确认为体外抑制
BMP-7表达最为有效的干扰序列。结论BMP-7-3 siRNA序列能够在体外高效能抑制BMP-7表达，故可作为一种实验研究技
术手段，用于探讨BMP-7的生物学作用机制的基础和临床应用研究。

Abstract: Objective To obtain highly efficient small interfering RNAs (siRNAs) against bone morphogenetic protein-7 (BMP-7)
for future functional analysis of BMP-7 in bone homeostasis. Methods We designed 4 independent siRNA sequences against
rat BMP-7 and transfected them in rat umbilical vein endothelial cells. After 72 h, we examined BMP-7 mRNA and protein
level in the transfected cells using real-time PCR and Western blotting, respectively. Results All the 4 siRNAs effectively
reduced BMP-7 expression in rat umbilical vein endothelial cells, and among them BMP-7-3 siRNA showed the highest
efficiency. Conclusion We have obtained an efficient siRNA to knockdown BMP-7 expression in vitro for further investigation
of the role of BMP-7 in bone and cartilage formation.