The transcription factor cyclic adenosine 3′,5′‐monophosphate response element‐binding protein enhances the odonto/osteogenic differentiation of stem cells from the apical papilla
The transcription factor cyclic adenosine 3′,5′‐monophosphate response element‐binding protein enhances the odonto/osteogenic differentiation of stem cells from the apical papilla
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DOI：
10.1111/iej.12709
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发表时间：
2017-09
影响因子：
5
通讯作者：
S. Su;Y. Zhu;S. Li;Y. Liang;J. Zhang
中科院分区：
文献类型：
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作者：
S. Su;Y. Zhu;S. Li;Y. Liang;J. Zhang
目的探讨cAMP反应元件结合蛋白（CREB）在根尖乳头（SCAPs）干细胞向牙状/成骨分化中的调控作用。方法从人阻生第三磨牙（n = 15）获得根尖乳头干细胞。用CREB过表达/沉默慢病毒转染分离的SCAPs。转染后的细胞用茜素红染色，观察矿化结节的形成。采用定量逆转录聚合酶链式反应（qRT-PCR）检测矿化相关基因碱性磷酸酶（ALP）、ⅰ型胶原蛋白（Col I）、矮子相关转录因子2 （RUNX2）、骨甾体（OSX）和骨钙素（OCN）的表达。Western blotting分析牙本质涎蛋白（DSP）和成骨相关标志物RUNX2的表达。统计学分析采用单因素方差分析（anova）和Student’st检验（a = 0.05）。结果过表达CREB可促进矿化结节形成，上调ALP、Col I、RUNX2、OSX、OCN等牙体/成骨相关标志物mRNA表达（P < 0.05），提高DSP、RUNX2蛋白表达（P < 0.05）。相比之下，CREB的沉默抑制了SCAPs的矿化能力（P < 0.05），降低了牙体/成骨相关标志物的表达（P < 0.05）。结论CREB表达的上调促进了SCAPs的牙髓/成骨分化，为牙本质-牙髓复合体的再生提供了一种可能的方法。
AIM To investigate the role of cAMP response element-binding protein (CREB) in the regulation of odonto/osteogenic differentiation of stem cells from the apical papilla (SCAPs). METHODOLOGY Stem cells from the apical papilla were obtained from human impacted third molars (n = 15). Isolated SCAPs were transfected with CREB overexpressing/silenced lentivirus. Transfected cells were stained with alizarin red to investigate mineralized nodule formation. The expression of the mineralization-related genes, alkaline phosphatase (ALP), collagen type I (Col I), runt-related transcription factor 2 (RUNX2), osterix (OSX) and osteocalcin (OCN), was determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Protein expression of the odontogenic-related marker dentine sialoprotein (DSP) and the osteogenic-related marker RUNX2 was measured by Western blotting analysis. One-way analysis of variance (anova) and Student's t-test were used for statistical analysis (a = 0.05). RESULTS The overexpression of CREB enhanced mineralized nodule formation and up-regulated (P < 0.05) the mRNA levels of odonto/osteogenic-related markers, including ALP, Col I, RUNX2, OSX and OCN, and also increased (P < 0.05) the protein expression of DSP and RUNX2. In contrast, the silencing of CREB inhibited (P < 0.05) the mineralization capacity of the SCAPs and decreased (P < 0.05) the expression of odonto/osteogenic-related markers. CONCLUSION Up-regulation of CREB expression promoted odonto/osteogenic differentiation of SCAPs and provided a potential method for the regeneration of the dentine-pulp complex.