Endothelial differentiation of adipose-derived mesenchymal stem cells is improved by epigenetic modifying drug BIX-01294
Endothelial differentiation of adipose-derived mesenchymal stem cells is improved by epigenetic modifying drug BIX-01294
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DOI：
10.1016/j.ejcb.2012.11.001
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发表时间：
2013-02-01
影响因子：
6.6
通讯作者：
Pelisek, Jaroslav
中科院分区：
文献类型：
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作者：
Culmes, Mihaela;Eckstein, Hans-Henning;Pelisek, Jaroslav
染色质重塑在基因转录调控中起着重要作用。因此，染色质的靶向性改变也可能增强体细胞的多能性。本研究旨在评价组蛋白G9a抑制剂表观遗传药物Bix-01294(Bix)对间充质干细胞DNA甲基化、多能基因Pou5f1(异构体a)、NANOG、KLF4和Cmyc表达的影响，以及提高其向内皮细胞分化的能力。从腹部脂肪组织中分离培养出人脂肪间充质干细胞(AdMSCs)。细胞用Bix预处理48h，在内皮细胞培养液中进一步分化7d和14d。通过甲基光的应用、多能性基因的表达、基于SYBRGreen的实时荧光聚合酶链式反应、免疫细胞化学和免疫印迹来确定全球DNA甲基化。经Bix处理后，AdMSCs的甲基化状态显著降低53%(p=0.008)，Pou5f1和NANOG的表达分别增加2.2倍(p=0.016)和1.5倍(p<；0.001)。此外，与未经Bix处理的AdMSCs相比，Bix预处理可提高AdMSCs向内皮细胞分化的能力，并显著增加VCAM-1、PECAM-1、von Willebrand因子、VEGFR-2、PDGF和Ang-1等血管内皮细胞标志物和血管生成因子的表达。在目前的研究中，我们证明了表观遗传修饰药物BIX-01294能够提高AdMSCs分化为ECs的能力，参与DNA和组蛋白甲基化。因此，Bix-01294可能是一种简单的提高AdMSCs分化潜能的工具。(C)2012年爱思唯尔股份有限公司。版权所有。
Chromatin remodeling plays an essential role in regulation of gene transcription. Consequently, targeted changes in chromatin may also augment pluripotency of somatic cells. The aim of the present study was to evaluate the effect of epigenetic drug BIX-01294 (BIX), a histone G9a inhibitor, on DNA methylation, expression of pluripotency genes POU5F1 (isoform a), NANOG, KLF4, and CMYC in mesenchymal stem cells, and the ability to increase their differentiation potential into endothelial cells (ECs). Human adipose-derived mesenchymal stem cells (AdMSCs) were isolated from abdominal adipose tissue. Cells were pre-treated with BIX for 48 h and further differentiated in endothelial medium for 7 and 14 days. Global DNA methylation was determined by MethyLight application, expression of genes for pluripotency, endothelial and angiogenic markers by SYBRGreen-based real-time PCR, immunocytochemistry, and immunobloting. Following treatment with BIX, DNA methylation status of AdMSCs was significantly reduced by 53% (p = 0.008), the expression of POU5F1 and NANOG was increased by 2.2-fold (p = 0.016) and 1.5-fold (p < 0.001), respectively. Furthermore, BIX pre-treatment improved the differentiation capacity of AdMSCs into ECs and significantly increased expression of several endothelial markers and factors involved in blood vessel formation: VCAM-1, PECAM-1, von Willebrand factor, VEGFR-2, PDGF, and ANG-1 in comparison with AdMSCs without BIX pre-treatment. In the present study we demonstrate that epigenetic modifying drug BIX-01294 is able to increase the ability of AdMSCs to differentiate into ECs engaging DNA and histone methylation. Hence, BIX-01294 might serve as a simple tool to increase the differentiation potential of AdMSCs. (c) 2012 Elsevier GmbH. All rights reserved.