miRNA-720 controls stem cell phenotype, proliferation and differentiation of human dental pulp cells.
miRNA-720 controls stem cell phenotype, proliferation and differentiation of human dental pulp cells.
复制标题
DOI：
10.1371/journal.pone.0083545
复制
发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Kuboki T
中科院分区：
文献类型：
--
作者：
Hara ES;Ono M;Eguchi T;Kubota S;Pham HT;Sonoyama W;Tajima S;Takigawa M;Calderwood SK;Kuboki T
牙髓细胞（DPC）富含干/祖细胞，但尚未得到很好的表征。小的非编码microRNA（miRNAs）被鉴定为控制蛋白质翻译、mRNA稳定性和转录，并且已经报道在干细胞生物学中发挥重要作用，与细胞重编程、维持干细胞性和调节细胞分化有关。为了表征牙髓干/祖细胞及其分化机制，我们在此从人DPC和牙周膜细胞（PDLC）中分选干细胞富集的侧群（SP）细胞，并进行基于锁核酸（LNA）的miRNA阵列。结果，与SP细胞相比，miR-720在分化的主群体（MP）细胞中高表达。计算机模拟分析和报告基因分析显示，miR-720靶向干细胞标志物NANOG，表明miR-720可通过抑制NANOG促进牙髓干/祖细胞分化。事实上，功能获得和丧失分析表明，miR-720控制NANOG转录和蛋白水平。此外，miR-720的转染显著减少了早期干细胞标志物SSEA-4阳性的细胞数量。同时，已知在干细胞分化过程中起关键作用的DNA甲基转移酶（DNMT）的mRNA水平也通过未知机制被miR-720增加。最后，miR-720通过针对ki-67的免疫细胞化学分析确定降低DPC增殖，并且通过茜素红染色以及碱性磷酸酶和骨桥蛋白mRNA水平证明促进牙源性分化。我们的研究结果确定miR-720是一种新的调控DPC分化的miRNA。
Dental pulp cells (DPCs) are known to be enriched in stem/progenitor cells but not well characterized yet. Small non-coding microRNAs (miRNAs) have been identified to control protein translation, mRNA stability and transcription, and have been reported to play important roles in stem cell biology, related to cell reprogramming, maintenance of stemness and regulation of cell differentiation. In order to characterize dental pulp stem/progenitor cells and its mechanism of differentiation, we herein sorted stem-cell-enriched side population (SP) cells from human DPCs and periodontal ligament cells (PDLCs), and performed a locked nucleic acid (LNA)-based miRNA array. As a result, miR-720 was highly expressed in the differentiated main population (MP) cells compared to that in SP cells. In silico analysis and a reporter assay showed that miR-720 targets the stem cell marker NANOG, indicating that miR-720 could promote differentiation of dental pulp stem/progenitor cells by repressing NANOG. Indeed, gain-and loss-of-function analyses showed that miR-720 controls NANOG transcript and protein levels. Moreover, transfection of miR-720 significantly decreased the number of cells positive for the early stem cell marker SSEA-4. Concomitantly, mRNA levels of DNA methyltransferases (DNMTs), which are known to play crucial factors during stem cell differentiation, were also increased by miR-720 through unknown mechanism. Finally, miR-720 decreased DPC proliferation as determined by immunocytochemical analysis against ki-67, and promoted odontogenic differentiation as demonstrated by alizarin red staining, as well as alkaline phosphatase and osteopontin mRNA levels. Our findings identify miR-720 as a novel miRNA regulating the differentiation of DPCs.
登录
查看更多内容
影响因子：
3.7
作者：
Eguchi T;Watanabe K;Hara ES;Ono M;Kuboki T;Calderwood SK
通讯作者：
Calderwood SK
影响因子：
64.8
作者：
Hatfield, SD;Shcherbata, HR;Ruohola-Baker, H
通讯作者：
Ruohola-Baker, H
DOI：
10.1073/pnas.0707594105
发表时间：
2008-02-05
影响因子：
11.1
作者：
Place, Robert F.;Li, Long-Cheng;Dahiya, Rajvir
通讯作者：
Dahiya, Rajvir
影响因子：
2.1
作者：
Iida, T;Suetake, I;Tsurimoto, T
通讯作者：
Tsurimoto, T
影响因子：
21.3
作者：
Chang, Chun-Ju;Chao, Chi-Hong;Xia, Weiya;Yang, Jer-Yen;Xiong, Yan;Li, Chia-Wei;Yu, Wen-Hsuan;Rehman, Sumaiyah K.;Hsu, Jennifer L.;Lee, Heng-Huan;Liu, Mo;Chen, Chun-Te;Yu, Dihua;Hung, Mien-Chie
通讯作者：
Hung, Mien-Chie