CAJ | 학술논문

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。
목적：탐토응용소간우RNA（ small interfering RNA，siRNA）특이성억제토골수간충질간세포내Runx2기인표체，사선고효특이성siRNA。방법근거siRNA설계원칙，침대Runx2기인서렬특정설계Runx2특이siRNA（1-3），전염토골수간충질간세포，용QPCR화Western blot방법검측siRNA대Runx2기인적억제효과。결과 Runx2 siRNA-2가유효억제골수간충질간세포중Runx2기인적표체。수siRNA-2종농도유50 nmol/L증가도100 nmol/L급200 nmol/L，억제효솔축점증강（ P＜0．05）；siRNA-2이종농도200 nmol/L전염후48 h억제효과최강，72 h축점감약，단잉명현억제（ P＜0．05）。결론응용RNA간우기술가억제골수간충질간세포중Runx2기인적표체，기억제작용구유명현적시간、농도의뢰성。
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.