Laboratory of Cellular and Molecular Engineering "S. Cavalcanti", University of Bologna, Campus of Cesena, via Venezia, 52, 47521, Cesena, FC, Italy.
Cell Biochem Biophys. 2013 Nov;67(2):255-62. doi: 10.1007/s12013-013-9610-z.
Adipose-derived stem cells (ADSCs) are stromal mesenchymal stem cells isolated from lipoaspirates, and they display a broad potential to differentiate toward different lineages. The role of epigenetics in regulating the expression of their lineage-specific genes is under evaluation, however till date virtually nothing is known about the relative significance of cardiac-specific transcription factor genes in human ADSCs. The aim of this study was to investigate DNA promoter methylation and relevant histone modifications involving MEF-2C, GATA-4, and Nkx2.5 in native human ADSCs. CpG sites at the transcription start in their promoters were found unmethylated using methylation-specific PCR. Chromatin immunoprecipitation assay showed low levels of total acetylated H3 histone (acH3) and high levels of trimethylated lysine 27 in H3 histone (H3K27me3) which were associated with both GATA-4 and Nkx2.5 promoters, indicating their transcriptional repressive chromatin arrangement. On the other hand, the opposite was apparent for MEF-2C promoter. Accordingly, MEF-2C-but not GATA-4 and Nkx2.5-transcripts were evidenced in native human ADSCs. These results suggest that the chromatin arrangement of these early cardiac regulatory genes could be explored as a level of intervention to address the differentiation of human ADSCs toward the cardiac lineage.
脂肪干细胞（ADSCs）是从脂肪抽吸物中分离出来的基质间充质干细胞，它们具有向不同谱系分化的广泛潜力。然而，表观遗传学在调节其谱系特异性基因表达中的作用仍在评估中，迄今为止，人们对人心肌 ADCS 中心脏特异性转录因子基因的相对重要性几乎一无所知。本研究旨在研究涉及 MEF-2C、GATA-4 和 Nkx2.5 的 DNA 启动子甲基化和相关组蛋白修饰在天然人心肌 ADCS 中的作用。使用甲基特异性 PCR 发现它们启动子转录起始处的 CpG 位点未甲基化。染色质免疫沉淀分析显示，与 GATA-4 和 Nkx2.5 启动子相关的总乙酰化 H3 组蛋白（acH3）水平较低，H3 组蛋白（H3K27me3）中赖氨酸 27 三甲基化水平较高，表明其转录抑制性染色质排列。另一方面，MEF-2C 启动子则相反。因此，在天然人心肌 ADCS 中可以检测到 MEF-2C-但不是 GATA-4 和 Nkx2.5-的转录物。这些结果表明，这些早期心脏调节基因的染色质排列可以作为一种干预水平来解决人心肌 ADCS 向心脏谱系的分化。
