摘要
目的探讨应用小干扰RNA(small interfering RNA,siRNA)特异性抑制兔骨髓间充质干细胞内Runx2基因表达,筛选高效特异性siRNA。方法根据siRNA设计原则,针对Runx2基因序列特征设计Runx2特异siRNA(1-3),转染兔骨髓间充质干细胞,用QPCR和Western blot方法检测siRNA对Runx2基因的抑制效果。结果 Runx2 siRNA-2可有效抑制骨髓间充质干细胞中Runx2基因的表达。随siRNA-2终浓度由50 nmol/L增加到100 nmol/L及200 nmol/L,抑制效率逐渐增强(P<0.05);siRNA-2以终浓度200 nmol/L转染后48 h抑制效果最强,72 h逐渐减弱,但仍明显抑制(P<0.05)。结论应用RNA干扰技术可抑制骨髓间充质干细胞中Runx2基因的表达,其抑制作用具有明显的时间、浓度依赖性。
Objective To investigate the inhibitory effect of Runx2 specific small interfering RNA （ siRNA） on Runx2 gene expression in rabbit bone marrow mesenchymal stem cells, and to screen the most efficient Runx2 specific siRNA.Methods According to the principle of siRNA design, Runx2 specific siRNA （1-3） was designed and the rabbit bone marrow mesenchymal stem cells were transfected.The expression level of Runx2 gene was detected using real-time quantitative PCR and Western blotting. Results Endogenous Runx2 expression was efficiently blocked in bone marrow mesenchymal stem cells by Runx2 siRNA-2 in a dose and time dependent manner.The expression of Runx2 decreased significantly along with the increase of the concentration of siRNA-2 from 50 nmol/L to 100 and 200 nmol/L.After 48h transfection with 200 nmol/L siRNA-2, the inhibitory effect was the strongest.It decreased after 72 hours, but the inhibition was still obvious （P〈0.05）.Conclusion The expression of Runx2 in bone marrow mesenchymal stem cells can be efficiently blocked in a dose and time dependent manner by specific siRNA.
出处
《中国骨质疏松杂志》
CAS
CSCD
北大核心
2014年第9期1089-1092,共4页
Chinese Journal of Osteoporosis
