口腔医学研究

口腔医学研究 ›› 2019, Vol. 35 ›› Issue (5): 488-492.DOI: 10.13701/j.cnki.kqyxyj.2019.05.018

• 口腔干细胞与口腔生物学研究 • 上一篇    下一篇

骨形态发生蛋白受体2在诱导多能干细胞成釉分化中的作用研究

刘治1*, 秦青2, 高鹏3, 孙聪1, 张乐琪1   

  1. 1. 西安交通大学第一附属医院口腔科 710000;
    2. 解放军空军第986医院口腔科 陕西 西安 710000;
    3. 新乡医学院第三附属医院口腔科 河南 新乡 453000
  • 收稿日期:2018-10-22 出版日期:2019-05-28 发布日期:2019-05-21
  • 通讯作者: 刘治,E-mail:kjiao1@163.com
  • 作者简介:刘治(1983~),男,汉族,陕西西安人,博士,主治医师,主要从事组织工程化牙齿研究。
  • 基金资助:
    西安交通大学第一附属医院创新项目(编号:2017011)

Effects of Bone Morphogenetic Protein Receptor 2 on Ameloblastic Differentiation of Induced Pluripotent Stem Cells.

LIU Zhi1*, QIN Qing2, GAO Peng3, SUN Cong1, ZHANG Yue-qi1   

  1. 1. Department of Stomatology, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710000, China;
    2. Department of Stomatology, The 986 Hospital of Air Force, Xi'an 710000, China;
    3. Department of Stomatology, the Thrid Hospital of Xinxiang Medical School, Xinxiang 453000, China.
  • Received:2018-10-22 Online:2019-05-28 Published:2019-05-21

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摘要： 目的: 研究骨形态发生蛋白受体2(bone morphogenetic protein receptor 2, BMPR-Ⅱ)在诱导多能干细胞(induced pluripotent stem,iPS)成釉分化中的作用。方法: 采用灭活小鼠胚胎成纤维细胞作为滋养层培养iPS,采用实时定量聚合酶链反应及Western blot检测成釉细胞无血清条件培养液(ameloblast serum-free conditioned medium,ASF-CM)或对照培养基培养iPS细胞14 d后,其BMPR-Ⅱ表达情况;采用BMPR-Ⅱ小干扰RNA(small interfering RNA, siRNA)或其阴性对照预处理iPS,再采用免疫荧光染色及流式细胞计数检测ASF-CM或对照培养基对iPS表达多种成釉标志蛋白的影响。结果: 经ASF-CM处理的iPS中BMPR-Ⅱ基因及蛋白表达水平均较对照组升高(P<0.05),ASF-CM培养组iPS中成釉蛋白、釉质素及细胞角蛋白-14表达水平均较对照组显著增高(P<0.05),BMPR-Ⅱ siRNA预处理可显著逆转ASF-CM促iPS成釉分化的作用(P<0.05)。结论: BMPR-Ⅱ信号转导在ASF-CM诱导的iPS成釉分化过程中发挥关键调控作用。

关键词: 骨形态发生蛋白受体2, 诱导多能干细胞, 成釉分化

Abstract: Objective: To investigate the effects of bone morphogenetic protein receptor 2 (BMPR-Ⅱ) on the ameloblastic differentiation of induced pluripotent stem cells (iPS). Methods: The mouse embryo fibroblasts treated with mitomycin-C were used as the feeder cells for iPS cells. Real-time PCR and western blot were used to measure the gene and protein expression of different BMP receptors by iPS cells cultured with control medium or ASF-CM. Immunohistochemical staining and flow cytometry were used to detect the expression of ameloblastin, enamelin, and cytokeratin-14 by BMPR-Ⅱ siRNA-pretreated iPS cells cultured with control medium or ASF-CM. Results: The gene and protein expression of BMPR-Ⅱ by iPS cells treated with ASF-CM were significantly higher than those of the controls (P<0.05). ASF-CM treatment significantly increased the gene and protein expression of ameloblastin, enamelin, and cytokeratin-14 (P<0.05), which could be obviously reversed by BMPR-Ⅱ siRNA pre-treatment (P<0.05). Conclusion: BMPR-Ⅱ plays an important role during the regulation of the ameloblastic differentiation of iPS cells.

Key words: Bone morphogenetic protein receptor 2, Induced pluripotent stem cells, Ameloblastic differentiation