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| 小鼠骨髓间充质干细胞Osterix基因特异siRNA的设计和沉默作用 |
| Design and silencing of Osterix specific siRNA in mouse bone marrow mesenchymal stem cells |
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| DOI：10.3969/j.issn.1006.7108.2015.12.010 |
| 中文关键词: RNA干扰 Osterix基因；小鼠骨髓间充质干细胞；基因沉默 |
| 英文关键词:RNA interference Osterix gene Mouse bone marrow mesenchymal stem cells Gene silencing |
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| 中文摘要: |
| 目的 探讨应用小干扰RNA(small interfering RNA，siRNA)特异性抑制骨髓间充质干细胞内Osterix基因表达，筛选高效特异性siRNA。方法 根据siRNA设计原则，针对Osterix基因序列特征设计Osterix特异siRNA( 1-3 )，转染骨髓间充质干细胞，用QPCR和Western blot方法检测siRNA对Oslerix基因的抑制效果。结果 Osterix siRNA-2可有效抑制骨髓间充质干细胞中Osterix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)；siHNA-2以终浓度200 rnnol/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Osterix基因的表达，其抑制作用具有明显的时间、浓度依赖性。 |
| 英文摘要: |
| Objective To study the inhibition effect of Osterix specific small interfering RNA (siRNA) on Osterix gene expression in bone marrow mesenchymal stem cells, and to screen the most efficient Osterix specific siRNA. Methods According to the principle of siRNA design, Osterix specific siRNA (1-3) was designed and the bone marrow mesenchymal stem cells were transfected. The expression of Osterix gene was detected using real-time quantitative PCR and Western blotting. Results Endogenous Osterix expression was efficiently blocked in bone marrow mesenchymal stem cells by Osterix siRNA-2 in a dose- and time-dependent manner. Osterix expression decreased significantly along with the increase of siRNA-2 dose within 50 - 200 nmol/L and it was the lowest at 48h after transfection with 200nmol/L of siRNA-2 (P< 0. 05). Conclusion The expression of Osterix in bone marrow mesenchymal stem cells can be blocked efficiently in a dose and time dependent manner by specific siRNA. |
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