慢病毒介导microRNA-214在成牙骨质细胞分化过程中的功能研究
The role of microRNA-214 in the differentiation and mineralization of cementoblasts
惠宏斌 1丁钰 1付建军 1屈小兵2
作者信息
- 1. 宝鸡市中医医院口腔科,陕西 宝鸡 721001
- 2. 宝鸡市第二中医医院口腔科, 陕西 宝鸡 721003
- 折叠
摘要
目的:构建microRNA-214沉默慢病毒载体并研究其在成牙骨质细胞分化过程中的作用.方法:常规条件下培养成牙骨质细胞系(OCCM-30 cell lines),并对其进行矿化诱导,检测牙骨质分化相关标记物的变化.构建microRNA-214沉默慢病毒载体并感染成牙骨质细胞后,用荧光显微镜检测其感染效率,cck8测定各组细胞的增殖速率;用茜素红和Von kossa染色检测各组成牙骨质细胞的矿化能力;Real-Time PCR检测各组细胞牙骨质分化标志物的表达水平.结果:常规诱导成牙骨质细胞并检测其标志物的改变,结果表明成牙骨质细胞矿化诱导成功.荧光显微镜显示,microRNA-214沉默载体能够顺利进入细胞;cck8测定结果显示,microRNA-214沉默组能显著提高成牙骨质细胞的增殖速率;茜素红和Von kossa染色结果发现,空白组和阴性对照组矿化结节形成量较少,而microRNA-214沉默组则形成较多的矿化结节;成牙骨质细胞矿化标志物检测显示,microRNA-214沉默组成牙骨质矿化标志物表达量较高(P<0.05).结论:microRNA-214沉默能够促进成牙骨质细胞的分化.
Abstract
AIM: To study the role of microRNA-214 in the differentiation and mineralization of cementoblasts.METHODS: Silencing lentiviral vector of microRNA-214 was constructed and then transfected into mouse cementoblasts OCCM-30 cell line.The transfection efficiency and silencing effects of the construct were observed by fluorescence microscope.The mineralization ability of cementoblasts was examined by Alizarin Red and Von kossa staining.The differentiation ability of cementoblasts was tested by Real-time PCR.RESULTS: The fluorescence microscope showed that the silencing lentiviral vector of microRNA-214 could be successfully transfected into cementoblasts.Differentiation of cementoblasts was demonstrated by the presence of the differentiation markers.The Alizarin Red and Von kossa staining demonstrated that the microRNA-214 silencing construct could boost the differentiation and minerlization ability of cementoblasts.CONCLUSION: Silencing of microRNA-214 can promote the differentiation and mineralization ability of cementoblasts.
关键词
microRNA-214/慢病毒/成牙骨质细胞/分化和矿化Key words
microRNA-214/lentiviral vector/cementoblasts/differentiation and mineralization引用本文复制引用
出版年
2017
NETL
NSTL
维普
万方数据