Runx2基因高表达对脐血间充质干细胞成骨分化相关基因表达的影响

发布时间：2018-03-18 23:35

本文选题：脐血干细胞移植　切入点：核心结合因子α亚基　出处：《中国组织工程研究》2017年09期 　论文类型：期刊论文

【摘要】：背景:Runx2在成骨分化的信号途径中起核心作用,在诱导干细胞成骨分化及促进骨愈合方面具有重要的研究意义。目的:观察Runx2基因高表达对脐血间充质干细胞成骨分化的影响。方法:(1)扩增Runx2基因,利用腺病毒载体构建Runx2重组腺病毒(pA d-Runx2),检测病毒滴度后用重组腺病毒感染脐血间充质干细胞,并于荧光显微镜下观察荧光表达变化;(2)感染后1,3,7,14 d采用实时荧光定量PCR及Western blot检测脐血间充质干细胞Runx2、BMP-2、OCN、ALP的mRNA和蛋白表达。结果与结论:(1)成功制备pA d-Runx2重组腺病毒,病毒滴度为1.7×10~(10)pfu/L;(2)Runx2重组腺病毒感染脐血间充质干细胞后细胞形态呈成骨样细胞改变,对照组细胞无明显变化;(3)感染组细胞Runx2、BMP-2、OCN、ALP mRNA和蛋白的表达量随感染时间的增加均有不同程度的上调;(4)结果表明,高表达Runx2基因腺病毒感染脐血间充质干细胞,能够上调BMP-2、OCN、ALP基因的表达,从而促进脐血间充质干细胞的成骨分化。
[Abstract]:Background: Runx2 plays a central role in the signaling pathway of osteogenic differentiation. It has important significance in inducing osteogenic differentiation of stem cells and promoting bone healing. Objective: to observe the effect of high expression of Runx2 gene on osteogenic differentiation of umbilical cord blood mesenchymal stem cells. Adenovirus vector was used to construct Runx2 recombinant adenovirus pAd-Runx2. After detecting the titer of the recombinant adenovirus, mesenchymal stem cells were infected with recombinant adenovirus, and the recombinant adenovirus was used to infect umbilical cord blood mesenchymal stem cells. The expression of mRNA and protein in umbilical cord blood mesenchymal stem cells Runx2BMP-2OCN-ALP was detected by real-time quantitative PCR and Western blot on the 14th day after infection. Results and conclusion the recombinant adenovirus pAd-Runx2 was successfully prepared. The titer of the virus was 1.7 脳 10 ~ (10) pfup / L ~ (2 +) rrrrrx2 recombinant adenovirus infected umbilical cord blood mesenchymal stem cells with osteoblast-like changes in morphology. The results showed that the expression of mRNA and protein of Runx2BMP-2OCNP in infected cells were up-regulated in varying degrees with the time of infection. The results showed that adenovirus with high expression of Runx2 gene was infected with mesenchymal stem cells of umbilical cord blood. It can up-regulate the expression of BMP-2 OCN-ALP gene and promote the osteogenic differentiation of umbilical cord blood mesenchymal stem cells.
【作者单位】：新乡市中心医院脊柱外科;郑州大学生命科学学院;郑州大学第一附属医院;
【基金】：国家自然科学基金资助项目(U1404313),课题名称:HDAC1双向调控在脐带MSCs向神经元样细胞定向分化过程中的作用和机制研究~~
【分类号】：R68

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