›› 2018, Vol. 38 ›› Issue (7): 582-586.

• 基础研究 • 上一篇    下一篇

CD24基因调控根尖牙乳头干细胞的成骨分化功能

王海锋1,刘亚男1,刘思思1,宁美芝1,幸丹1,李文峰1,曹钰2   

  1. 1. 首都医科大学康复医学院,中国康复研究中心北京博爱医院北京
    2. 首都医科大学附属北京口腔医院
  • 收稿日期:2018-01-12 修回日期:2018-03-07 出版日期:2018-07-28 发布日期:2018-07-28
  • 通讯作者: 曹钰 E-mail:caoyu_bj@163.com
  • 基金资助:
    国家自然科学基金面上项目;中国康复研究中心重点项目

CD24 mediate the osteogenic differentiation potential of stem cells from apical papilla

  • Received:2018-01-12 Revised:2018-03-07 Online:2018-07-28 Published:2018-07-28

摘要: 目的 研究干细胞表面标记CD24基因对根尖牙乳头干细胞成骨分化能力的影响。方法 利用CD24 shRNA基因敲除CD24进行丧失性功能研究;实时定量RT-PCR检测CD24的基因敲除效果;通过ALP活性检测、茜素红染色及钙离子定量分析明确根尖牙乳头干细胞体外成骨分化能力的变化;实时荧光定量RT-PCR检测成骨分化相关基因-I型胶原蛋白1A、I型胶原蛋白1B、骨涎蛋白、骨桥蛋白和成骨相关转录因子RUNX2的表达分析研究根尖牙乳头干细胞基因表达改变。结果 实时定量RT-PCR结果显示CD24可以在根尖牙乳头干细胞有效的被敲除;碱性磷酸酶活性结果显示敲除CD24抑制根尖牙乳头干细胞碱性磷酸酶活性;茜素红染色及钙离子定量分析结果显示敲除CD24明显抑制根尖牙乳头干细胞体外矿化能力;实时定量RT-PCR结果显示敲除CD24明显抑制I型胶原蛋白A、型胶原蛋白B、骨涎蛋白、骨桥蛋白和RUNX2的表达。结论 基因沉默CD24可以通过抑制转录因子RUNX2及成骨分化基因的表达,从而抑制根尖牙乳头干细胞体外成骨分化功能。

关键词: CD24, 根尖牙乳头干细胞, 成骨分化

Abstract: Abstract:Objective To investigate the effect of CD24 molecule(CD24) on the osteogenic differentiation potential of stem cells from apical papilla (SCAPs). Methods Lentiviral CD24 ShRNA was used to silence the expression of CD24. Real-time fluorescence quantitative RT-PCR was used to detect the knockout effect of CD24. Alkaline phosphatase (ALP) activity assay was used to detect the ALP activity, which was the early marker of osteogenic differentiation. Alizarin-red staining and calcium quantitative analysis were used to study the potential of SCAPs mineralization in vitro. The expression of osteogenic related genes was detected by real-time fluorescence quantitative RT-PCR, including collagen type I alpha 1 chain (COL1A1), collagen type I alpha 2 chain (COL1A2), bone sialoprotein (BSP), osteonectin (ON) and runt-related transcription factor 2 (RUNX2). The gene expression changes of SCAPs were analyzed. Results Real time RT-PCR result showed that CD24 could be silenced in SCAPs. ALP activity assay showed that the depletion of CD24 inhibited ALP activity. Alizarin-red staining and calcium quantitative analysis revealed that the knockout of CD24 obviously suppressed the mineralization of SCAPs in vitro. Real-time fluorescence quantitative RT-PCR displayed that the depletion of CD24 obviously depressed the expressions of COL1A1, COL1A2, BSP and ON in SCAPs. The RUNX2 expression was repressed in CD24 silenced SCAPs compared with control group. Conclusion The gene silencing of CD24 represses the osteogenic differentiation potential of SCAPs by targeting the RUNX2.

Key words: CD24 molecule (CD24), Stem cells from apical papilla (SCAPs), Osteogenic differentiation