Xu Rui, Chen Wenbin, Zhang Zhifen, Qiu Yang, Wang Yong, Zhang Bingchang, Lu Wei
Department of Clinical Laboratory Medicine, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, Shandong 250021, China.
Central Laboratory, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, Shandong 250021, China.
Gene. 2018 May 30;657:9-18. doi: 10.1016/j.gene.2018.03.010. Epub 2018 Mar 5.
Bone-Marrow Stromal Cells (BMSCs)-derived vascular endothelial cells (VECs) is regarded as an important therapeutic strategy for spinal cord injury, disc degeneration, cerebral ischemic disease and diabetes. The change in DNA methylation level is essential for stem cell differentiation. However, the DNA methylation related mechanisms underlying the endothelial differentiation of BMSCs are not well understood. In this study, DNA methyltransferase inhibitor, 5-aza-2'-deoxycytidine (5-aza-dC) significantly elevated the endothelial markers expression (CD31/PECAM1, CD105/ENG, eNOS and VE-cadherin), as well as promoted the capacity of angiogenesis on Matrigel. The result of Alexa 488-Ac-LDL uptake assay indicated that the differentiation ratio of BMSCs into VECs was 68.7% in 5-azaz-dC induced differentiation. And then we screened differentiation inducers with altered expression patterns and DNA methylation levels in four important families (VEGF, ANG, FGF and ETS). By integrating these data, five endothelial differentiation inducers (VEGFA, ANGPT2, FGF2, FGF9 and ETS1) which were directly upregulated by 5-aza-dC and five indirect factors (FGF1, FGF3, ETS2, ETV1 and ETV4) were identified. These data suggested that 5-aza-dC is an excellent chemical molecule for BMSCs differentiation into functional VECs and also provided essential clues for DNA methylation related signaling during 5-aza-dC induced endothelial differentiation of BMSCs.
骨髓基质细胞（BMSCs）衍生的血管内皮细胞（VECs）被视为脊髓损伤、椎间盘退变、脑缺血疾病和糖尿病的一种重要治疗策略。DNA甲基化水平的变化对干细胞分化至关重要。然而，BMSCs向内皮细胞分化的DNA甲基化相关机制尚不清楚。在本研究中，DNA甲基转移酶抑制剂5-氮杂-2'-脱氧胞苷（5-aza-dC）显著提高了内皮细胞标志物的表达（CD31/PECAM1、CD105/ENG、eNOS和VE-钙黏蛋白），同时促进了基质胶上的血管生成能力。Alexa 488-Ac-LDL摄取试验结果表明，在5-氮杂-dC诱导分化中，BMSCs向VECs的分化率为68.7%。然后，我们在四个重要家族（VEGF、ANG、FGF和ETS）中筛选了表达模式和DNA甲基化水平发生改变的分化诱导剂。通过整合这些数据，鉴定出了五个由5-aza-dC直接上调的内皮细胞分化诱导剂（VEGFA、ANGPT2、FGF2、FGF9和ETS1）和五个间接因子（FGF1、FGF3、ETS2、ETV1和ETV4）。这些数据表明，5-aza-dC是一种用于BMSCs分化为功能性VECs的优良化学分子，也为5-aza-dC诱导BMSCs内皮细胞分化过程中与DNA甲基化相关的信号传导提供了重要线索。
