›› 2017, Vol. 37 ›› Issue (5): 413-417.

• 基础研究 • 上一篇    下一篇

Rap1b对鼠前成骨细胞Mc3T3-E1成骨分化早期的影响

施歌1,马俊青1,赵超越2,陈行素3,朱零4,陈文静1   

  1. 1. 江苏省口腔医院
    2. 南京医科大学口腔医学院
    3. 南京医科大学附属口腔医院正畸科
    4. 南京医科大学口腔疾病研究江苏省重点实验室,南京医科大学附属口腔医院
  • 收稿日期:2016-11-20 修回日期:2017-01-19 出版日期:2017-05-28 发布日期:2017-05-19
  • 通讯作者: 陈文静 E-mail:chenwenjing_orth@163.com

Effect of Rap1b on early osteogenic differentiation of mouse preosteoblasts (Mc3T3-E1)

  • Received:2016-11-20 Revised:2017-01-19 Online:2017-05-28 Published:2017-05-19

摘要: 目的 探讨Rap1b对鼠前成骨细胞Mc3T3-E1成骨分化早期的影响。 方法 利用成骨诱导液诱导Mc3T3-E1成骨向分化,利用实时定量PCR分别检测诱导后0、3、7 d的Rap1b mRNA的相对表达量。同时向实验组细胞分别转染Rap1b-mus-462和Rap1b-mus-703,4 d后进行ALP染色,同时测定Rap1b的mRNA相对表达量,并对转染后细胞中成骨指标ALP、Runx2和Osterix 0、3、7 d时的mRNA相对表达量进行测定。再利用划痕实验观察siRNA转染对Mc3T3-E1细胞迁移能力的影响。 结果 Real-time PCR结果显示在成骨诱导早期的不同时间点,Rap1b的表达量随诱导时间的增加而增加。当Rap1b的表达降低时,各成骨指标ALP、Runx2和Osterix的表达量也随之降低,Mc3T3-E1细胞迁移能力也有所减弱。 结论 Rap1b对Mc3T3-E1细胞成骨分化早期以及细胞迁移有一定的促进作用。

关键词: Rap1b, 鼠前成骨细胞, mRNA相对表达量, 实时定量PCR

Abstract: Objective To investigate the effects of Rap1b on the early osteogenic differentiation of mouse preosteoblasts(Mc3T3-E1). Methods Mc3T3-E1 osteoblast differentiation was induced by osteogenic induction. The relative expression of mRNA Rap1b was detected by real-time quantitative PCR, at 0, 3 and 7 d respectively. At the same time, Rap1b-mus-462 and Rap1b-mus-703 were transfected to the experimental group cells respectively, and stained by ALP at 4 d. Then the mRNA relative expression of Rap1b was determined, and the mRNA expressions of ALP, Runx2 and Osterix at 0, 3, 7 d were also measured. And the effect of siRNA transfection on the migration of Mc3T3-E1 cells was observed by scratch test. Results The expression of Rap1b increased with the induction time in the early stage of osteogenic induction. The expression of Rap1b decreased after siRNA transfection, and the expression of Osterix, Runx2 and ALP decreased. The migration ability of Mc3T3-E1 cells decreased. Conclusion Rap1b can promote the osteogenic differentiation of Mc3T3-E1 cells and cell migration in the early stage.

Key words: Rap1b, rat osteoblast, mRNA relative expression, RT-q PCR

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