The epigenetic modifiers 5-aza-2'-deoxycytidine and trichostatin A influence adipocyte differentiation in human mesenchymal stem cells.
The epigenetic modifiers 5-aza-2'-deoxycytidine and trichostatin A influence adipocyte differentiation in human mesenchymal stem cells.
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DOI：
10.1590/1414-431x20132893
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发表时间：
2013-05
期刊：
影响因子：
--
通讯作者：
Correa A
中科院分区：
文献类型：
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作者：
Zych J;Stimamiglio MA;Senegaglia AC;Brofman PR;Dallagiovanna B;Goldenberg S;Correa A
表观遗传机制如DNA甲基化和组蛋白修饰在干细胞分化中是重要的。甲基化主要与转录抑制相关，而组蛋白乙酰化则与活跃的染色质状态相关。我们使用染色质修饰剂trichostatin A（一种组蛋白去乙酰化酶抑制剂）和5-aza-2 ' -脱氧胞苷（5azadC）（一种去甲基化剂）确定了这些表观遗传机制对骨髓间充质干细胞（MSCs）和脂肪组织（ADSCs）中脂肪细胞分化的影响。在脂肪形成开始前，用5、50或500 nM TSA或1、10或100µM 5azadC处理亚融合MSC培养2天。定量分化，评价脂肪细胞基因PPARG、FABP4及抗脂肪细胞基因GATA2的表达。除5 nM TSA处理的BM-MSCs外，TSA可减少脂肪生成。仅500 nM TSA处理可降低细胞增殖。5azadC处理在所有评估的条件下都降低了增殖和脂肪细胞分化，导致PPARG和FABP4下调，GATA2上调。ADSCs对治疗的反应强于BM-MSCs，这表明不同来源的细胞之间可能存在表观遗传记忆的差异。由于表观遗传特征影响分化，应该有可能通过考虑各种可用的来源来指导MSCs在细胞治疗中的使用，以提高过程效率。
Epigenetic mechanisms such as DNA methylation and histone modification are important in stem cell differentiation. Methylation is principally associated with transcriptional repression, and histone acetylation is correlated with an active chromatin state. We determined the effects of these epigenetic mechanisms on adipocyte differentiation in mesenchymal stem cells (MSCs) derived from bone marrow (BM-MSCs) and adipose tissue (ADSCs) using the chromatin-modifying agents trichostatin A (TSA), a histone deacetylase inhibitor, and 5-aza-2′-deoxycytidine (5azadC), a demethylating agent. Subconfluent MSC cultures were treated with 5, 50, or 500 nM TSA or with 1, 10, or 100 µM 5azadC for 2 days before the initiation of adipogenesis. The differentiation was quantified and expression of the adipocyte genes PPARG and FABP4 and of the anti-adipocyte gene GATA2 was evaluated. TSA decreased adipogenesis, except in BM-MSCs treated with 5 nM TSA. Only treatment with 500 nM TSA decreased cell proliferation. 5azadC treatment decreased proliferation and adipocyte differentiation in all conditions evaluated, resulting in the downregulation of PPARG and FABP4 and the upregulation of GATA2. The response to treatment was stronger in ADSCs than in BM-MSCs, suggesting that epigenetic memories may differ between cells of different origins. As epigenetic signatures affect differentiation, it should be possible to direct the use of MSCs in cell therapies to improve process efficiency by considering the various sources available.