ZBTB16 as a Downstream Target Gene of Osterix Regulates Osteoblastogenesis of Human Multipotent Mesenchymal Stromal Cells.
ZBTB16 as a Downstream Target Gene of Osterix Regulates Osteoblastogenesis of Human Multipotent Mesenchymal Stromal Cells.
复制标题
DOI：
10.1002/jcb.25634
复制
发表时间：
2016-10
影响因子：
4
通讯作者：
Izumi, Yuichi
中科院分区：
文献类型：
--
作者：
Onizuka, Satoru;Iwata, Takanori;Park, Sung-Joon;Nakai, Kenta;Yamato, Masayuki;Okano, Teruo;Izumi, Yuichi
关键词：
人多能间充质基质细胞(HMSCs)具有向成骨细胞分化的能力，可作为骨再生治疗的来源。在细胞移植前的骨组织工程中，通过诱导hMSCs在体外向成骨细胞分化的成骨诱导预处理已被广泛应用。然而，骨诱导介质(OIM)诱导成骨细胞分化的分子基础尚不清楚。因此，我们使用下一代测序仪来研究hMSCs在成骨分化过程中基因表达的变化。本研究所用的人骨髓间充质干细胞具有多能性和自我更新能力。全转录组分析显示，在hMSCs成骨过程中，锌指和含16个BTB结构域(ZBTB16)的表达显著增加。人骨髓间充质干细胞与OIM共同培养后，ZBTB16基因和蛋白的表达均增强。小干扰RNA(SiRNA)介导的ZBTB16基因沉默降低了碱性磷酸酶(ALP)的活性、骨钙素(OCN)和骨涎蛋白(BSP)等成骨基因的表达，以及OIM诱导的矿化结节形成。SiRNA介导的Osterix基因沉默(OSX)被认为是成骨细胞分化的重要调节因子，显著下调ZBTB16的表达。此外，染色质免疫沉淀(ChIP)分析表明，OSX与ZBTB16启动子区域结合，该区域包含富含GC的规范Sp1序列，这是OSX的特异性结合部位。这些发现表明，ZBTB16作为OSX的下游转录调节因子，可以作为成骨细胞分化的晚期标志物。J.细胞。生物化学。117：2423-2434,2016。©2016作者。《细胞生物化学杂志》由威利期刊出版公司出版。
Human multipotent mesenchymal stromal cells (hMSCs) possess the ability to differentiate into osteoblasts, and they can be utilized as a source for bone regenerative therapy. Osteoinductive pretreatment, which induces the osteoblastic differentiation of hMSCs in vitro, has been widely used for bone tissue engineering prior to cell transplantation. However, the molecular basis of osteoblastic differentiation induced by osteoinductive medium (OIM) is still unknown. Therefore, we used a next‐generation sequencer to investigate the changes in gene expression during the osteoblastic differentiation of hMSCs. The hMSCs used in this study possessed both multipotency and self‐renewal ability. Whole‐transcriptome analysis revealed that the expression of zinc finger and BTB domain containing 16 (ZBTB16) was significantly increased during the osteoblastogenesis of hMSCs. ZBTB16 mRNA and protein expression was enhanced by culturing the hMSCs with OIM. Small interfering RNA (siRNA)‐mediated gene silencing of ZBTB16 decreased the activity of alkaline phosphatase (ALP); the expression of osteogenic genes, such as osteocalcin (OCN) and bone sialoprotein (BSP), and the mineralized nodule formation induced by OIM. siRNA‐mediated gene silencing of Osterix (Osx), which is known as an essential regulator of osteoblastic differentiation, markedly downregulated the expression of ZBTB16. In addition, chromatin immunoprecipitation (ChIP) assays showed that Osx associated with the ZBTB16 promoter region containing the GC‐rich canonical Sp1 sequence, which is the specific Osx binding site. These findings suggest that ZBTB16 acts as a downstream transcriptional regulator of Osx and can be useful as a late marker of osteoblastic differentiation. J. Cell. Biochem. 117: 2423–2434, 2016. © 2016 The Authors. Journal of Cellular Biochemistry published by Wiley Periodicals, Inc.
登录
查看更多内容
影响因子：
4.8
作者：
Jose Ortuno, Maria;Ruiz-Gaspa, Silvia;Ventura, Francesc
通讯作者：
Ventura, Francesc
影响因子：
23.9
作者：
Caplan AI;Correa D
通讯作者：
Correa D
影响因子：
30.8
作者：
Barna, M;Hawe, N;Pandolfi, PP
通讯作者：
Pandolfi, PP
影响因子：
4.8
作者：
Matsubara, Takuma;Kida, Kumiko;Yoneda, Toshiyuki
通讯作者：
Yoneda, Toshiyuki
影响因子：
11.8
作者：
Bialek, P;Kern, B;Karsenty, G
通讯作者：
Karsenty, G