The lncRNA HOTAIR impacts on mesenchymal stem cells via triple helix formation.
The lncRNA HOTAIR impacts on mesenchymal stem cells via triple helix formation.
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DOI：
10.1093/nar/gkw802
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发表时间：
2016-12-15
影响因子：
14.9
通讯作者：
Wagner W
中科院分区：
文献类型：
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作者：
Kalwa M;Hänzelmann S;Otto S;Kuo CC;Franzen J;Joussen S;Fernandez-Rebollo E;Rath B;Koch C;Hofmann A;Lee SH;Teschendorff AE;Denecke B;Lin Q;Widschwendter M;Weinhold E;Costa IG;Wagner W
越来越多的人认为长链非编码RNA（lncRNA）调节细胞功能。在这项研究中，我们分析了lncRNA HOTAIR在间充质干细胞（MSC）中的作用，特别关注基因表达和DNA甲基化（DNAm）的衰老相关变化。HOTAIR结合位点在基因组区域富集，随着细胞培养物传代次数的增加而变得高度甲基化。HOTAIR的过表达和敲低分别抑制或刺激MSC的成脂分化。HOTAIR表达的修饰对基因表达仅引起非常温和的影响，特别是polycomb组靶基因。此外，HOTAIR的过表达和敲低导致HOTAIR结合位点的DNAm变化。根据反向Hoogsteen氢键，在HOTAIR序列内预测了五个潜在的三螺旋形成结构域。值得注意的是，这些HOTAIR结构域的预测的三螺旋靶位点也富集在差异表达的基因中，并且接近HOTAIR调节后的DNAm变化。电泳迁移率变动分析提供了HOTAIR结构域与预测的靶位点形成RNA-DNA-DNA三链体的进一步证据。我们的研究结果表明，HOTAIR影响MSC的分化，并且它与衰老相关的DNAm相关。将表观遗传修饰剂靶向基因组中的相关基因座可能涉及HOTAIR的三螺旋形成。
There is a growing perception that long non-coding RNAs (lncRNAs) modulate cellular function. In this study, we analyzed the role of the lncRNA HOTAIR in mesenchymal stem cells (MSCs) with particular focus on senescence-associated changes in gene expression and DNA-methylation (DNAm). HOTAIR binding sites were enriched at genomic regions that become hypermethylated with increasing cell culture passage. Overexpression and knockdown of HOTAIR inhibited or stimulated adipogenic differentiation of MSCs, respectively. Modification of HOTAIR expression evoked only very moderate effects on gene expression, particularly of polycomb group target genes. Furthermore, overexpression and knockdown of HOTAIR resulted in DNAm changes at HOTAIR binding sites. Five potential triple helix forming domains were predicted within the HOTAIR sequence based on reverse Hoogsteen hydrogen bonds. Notably, the predicted triple helix target sites for these HOTAIR domains were also enriched in differentially expressed genes and close to DNAm changes upon modulation of HOTAIR. Electrophoretic mobility shift assays provided further evidence that HOTAIR domains form RNA–DNA–DNA triplexes with predicted target sites. Our results demonstrate that HOTAIR impacts on differentiation of MSCs and that it is associated with senescence-associated DNAm. Targeting of epigenetic modifiers to relevant loci in the genome may involve triple helix formation with HOTAIR.