Long non-coding RNA H19 promotes the osteogenic differentiation of rat ectomesenchymal stem cells via Wnt/β-catenin signaling pathway
Long non-coding RNA H19 promotes the osteogenic differentiation of rat ectomesenchymal stem cells via Wnt/β-catenin signaling pathway
复制标题
DOI：
10.26355/eurrev_201812_16648
复制
发表时间：
2018-12-01
影响因子：
3.3
通讯作者：
Yang, Y-F
中科院分区：
文献类型：
--
作者：
Gong, Y-Y;Peng, M-Y;Yang, Y-F
目的：为探讨长链非编码RNA（In-cRNA）H19促进大鼠外胚间充质干细胞（EMSC）成骨分化的分子机制，采用流式细胞仪分离大鼠胚胎面突EMSC。荧光免疫法检测成骨标志物CD 29、CD 90、CD 44、CD 57、Nestin和sox 10。构建瞬时干扰H19、稳定表达H19的EMSCs细胞系及诱导成骨分化后，采用实时荧光定量PCR（RT-PCR）和Western blot检测β-catenin和Wnt通路靶基因的表达。用Wnt/β-连环蛋白信号通路抑制剂Wnt-C59处理H19过表达的EMSC，通过RT-PCR和Western印迹检测β-连环蛋白和成骨标志物的表达。结果：成功分离并鉴定了EMSC，其成骨标志物为CD 29、CD 90、CD 44、CD 57。Nestin和sox 10的表达量显著增加。成骨诱导液显著增加了EMSC中H19和成骨标志物ALP、Runx 2、BMP和OCN的表达（p
OBJECTIVE: To investigate the molecular mechanism of long noncoding RNA (In-cRNA) H19 that promotes osteogenic differentiation in rat ectomesenchymal stem cells (EMSCs).MATERIALS AND METHODS: EMSCs were isolated from rat fetal facial processes by flow cytometry. Osteogenic markers CD29, CD90, CD44, CD57, Nestin and sox10 were detected by fluorescent immunoassay. beta-catenin and Wnt pathway target genes were detected by Real-time polymerase chain reaction (RT-PCR) and Western blot after the construction of transient interference H19, a stable expression of H19 EMSCs cell line and the induction of osteogenic differentiation of EMSCs cells. EMSCs of H19 over-expression were treated with Wnt/beta-catenin signaling pathway inhibitor Wnt-C59, and the expressions of beta-catenin and osteogenic markers were detected by RT-PCR and Western blot. Furthermore, the mechanism of H19 regulating Wnt/beta-catenin signaling pathway was explored by transfecting miR-22 and miR-141 mimics and luciferase reporter assays.RESULTS: EMSCs were successfully isolated and identified, osteogenic markers CD29, CD90, CD44, CD57. Nestin and sox10 were significantly overexpressed. Osteogenesis-induced solution significantly increased the expression of H19 and osteogenic markers ALP, Runx2, BMP and OCN in EMSCs (p