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。
Objective To investigate the effects of Raplb 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 Raplb was detected by real-time quantitative PCR, at 0, 3 and 7 d respectively. At the same time, Raplb-mus-462 and Raplb-mus-703 were transfected to the experimental group cells respectively, and stained by ALP at 4 d. Then the mRNA relative expression of Raplb was determined, and the mRNA expressions of ALP, Runx2 and Osterix at 0, 3, 7 d were also measured. And the effect of siRNA transfec- tion on the migration of Mc3T3-E1 cells was observed by scratch test. Results The expression of Raplb increased with the induction time in the early stage of osteogenic induction. The expression of Raplb decreased after siRNA transfection, and the expression of Os- terix, Runx2 and ALP decreased. The migration ability of Mc3T3-EI cells decreased. Conclusion Raplb can promote the osteogenic differentiation of Mc3T3-EI cells and cell migration in the early stare.
出处
《口腔医学》
CAS
2017年第5期413-417,共5页
Stomatology
基金
优势学科二期项目(JX10531802/018)
