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-ⅡsiRNA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。
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-ⅡsiRNA-pretreated iPS cells cultured with control medium or ASF-CM.Results:The gene and protein expression of BMPR-Ⅱby 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-ⅡsiRNA pre-treatment(P<0.05).Conclusion:BMPR-Ⅱplays an important role during the regulation of the ameloblastic differentiation of iPS cells.
作者
刘治
秦青
高鹏
孙聪
张乐琪
LIU Zhi;QIN Qing;GAO Peng;SUN Cong;ZHANG Yue-qi(Department of Stomatology,The First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710000,China;Department of Stomatology,The 986 Hospital of Air Force,Xi'an 710000,China;Department of Stomatology,the Thrid Hospital of Xinxiang Medical School,Xinxiang 453000,China.)
出处
《口腔医学研究》
CAS
北大核心
2019年第5期488-492,共5页
Journal of Oral Science Research
基金
西安交通大学第一附属医院创新项目(编号:2017011)
关键词
骨形态发生蛋白受体2
诱导多能干细胞
成釉分化
Bone morphogenetic protein receptor 2
Induced pluripotent stem cells
Ameloblastic differentiation
