人根尖牙乳头干细胞与非干细胞差异表达miRNAs谱及靶基因的功能分析
Differential expression of target genes and gene function of miRNAs in human stem cells and non-stem cells from apical papilla
林诗晗 1胡雪刚 1吕红兵1
作者信息
- 1. 福建医科大学附属口腔医院牙体牙髓科, 福建 福州 350002
- 折叠
摘要
目的:比较人根尖牙乳头干细胞(SCAP)与非干细胞(非SCAP)之间差异表达的miRNAs,并进行功能预测分析.方法:采用酶消化法获得原代人根尖牙乳头细胞,用鼠抗人STRO-1单克隆抗体对其标记,用免疫磁珠分选系统获得SCAP和非SCAP.茜素红、油红O染色鉴定干细胞的分化能力;采用二代测序技术,检测SCAP和非SCAP中miRNAs的表达谱,并筛选差异表达(相差倍数>1.5)的miRNAs,用生物信息学方法对差异表达的miRNAs进行靶基因功能分析.结果:与非SCAP相比,SCAP中表达下调的miRNAs有has-miR-4532、has-miR-6131、has-miR-4284、has-miR-3182、has-miR-1273e、has-miR-7-1-3p、has-miR-31-3p、has-miR-3654、has-miR-495-3p、has-miR-382-3p,表达上调的miRNAs有has-miR-4508、has-miR-1247-5p、has-miR-6819-3p、has-miR-2116-3p、has-miR-490-5p、has-miR-335-5p、has-miR-1914-5p、has-miR-1249、has-miR-3180-3p、has-miR-668-3p.表达下调miRNAs所调控的下游靶基因主要有20个,表达上调的有6个,这些靶基因主要参与了细胞的自我更新、分化和迁移等生命过程.结论:SCAP特异性表达的miRNAs主要调控的下游靶基因及其基因功能可能与干性维持密切相关.
Abstract
AIM: To study the differential expression of target genes and gene function of miRNAs in human stem cells and non-stem cells from apical papilla (SCAPs and NSCPs).METHODS: SCAPs were isolated by immune-magnetic binding specific STRO-1 antibody separation system.Osteogenic and adipogenic differentiation of SCAPs were tested by alizarin red staining(ARS) and Oil Red O staining.Differential miRNAs of SCAPs and NSCPs were screened by the Next generation sequencing.Target genes and their possible roles were predicted using biological information.RESULTS: The results revealed that 10 miRNAs (hsa-miR-4532, hsa-miR-6131, hsa-miR-4284, hsa-miR-3182, hsa-miR-1273e, hsa-miR-7-1-3p, hsa-miR-31-3p, hsa-miR-3654, hsa-miR-495-3p, hsa-miR-382-3p) were downregulated while 10 miRNAs (hsa-miR-4508, hsa-miR-1247-5p, hsa-miR-6819-3p, hsa-miR-2116-3p, hsa-miR-490-5p, hsa-miR-335-5p, hsa-miR-1914-5p, hsa-miR-1249, hsa-miR-3180-3p, hsa-miR-668-3p) were upregulated in SCAPs compared with those of NSCPs (P<0.05).26 target genes which mainly involved in the self-renewal capability, cell differentiation and migration were found.CONCLUSION: The target genes and gene function regulated to specific miRNAs of SCAPs are closely related to the cell stemness.
关键词
根尖牙乳头干细胞/miRNAs/靶基因/基因功能/干性Key words
stem cells from the apical papilla (SCAPs)/miRNAs/targets genes/gene function/stemness引用本文复制引用
基金项目
福建省卫生系统中青年骨干人才培养项目(2014-ZQN-ZD-22)
出版年
2017
NETL
NSTL
维普
万方数据