Abstract

Homeobox transcript antisense RNA (HOTAIR) is a long non-coding RNA associated with a number of fibrosis-related diseases. The aim of this study was to investigate the specific role of HOTAIR in the development of endometrial fibrosis and to identify the molecular mechanisms underlying this process. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to determine the expression levels of HOTAIR in samples of intrauterine adhesion (IUA) tissue and in endometrial stromal cells (ESCs) that had been treated with transforming growth factor beta 1 (TGF-β1). Additionally, we transfected ESCs with either overexpression plasmid (pcDNA-HOTAIR) or silencing construct (si-HOTAIR) and then treated these cells with TGF-β1. We then performed RT-qPCR and western blot analysis, along with cell proliferation and apoptosis assays, to investigate the effects of HOTAIR on the transdifferentiation of ESCs into myofibroblasts. The results showed that the expression levels of HOTAIR were significantly elevated in IUA tissue and in ESCs that had been treated with TGF-β1. The overexpression of HOTAIR had a pro-fibrotic effect on ESCs, while the silencing of HOTAIR exerted an anti-fibrotic effect. Most importantly, the protein expression levels of p-Smad2 and p-Smad3 were significantly upregulated in TGF-β1-treated ESCs transfected with pcDNA-HOTAIR and were downregulated after transfection with si-HOTAIR constructs. These data indicate that HOTAIR promotes endometrial fibrosis by activating the TGF-β1/Smad signaling pathway, suggesting that the inhibition of HOTAIR may represent a promising therapeutic option for suppressing endometrial fibrosis.

中文翻译：

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LncRNA HOTAIR通过激活TGF-β1/ Smad途径促进子宫内膜纤维化

摘要

同源盒转录本反义RNA（HOTAIR）是一种长链非编码RNA，与多种纤维化相关疾病有关。这项研究的目的是调查HOTAIR在子宫内膜纤维化发展中的特定作用，并确定这一过程的分子机制。逆转录定量聚合酶链反应（RT-qPCR）用于确定子宫内粘连（IUA）组织样品和经转化生长因子β1（TGF）处理的子宫内膜基质细胞（ESC）中HOTAIR的表达水平-β1）。此外，我们用过表达质粒（pcDNA-HOTAIR）或沉默构建体（si-HOTAIR）转染ESC，然后用TGF-β1处理这些细胞。然后，我们进行了RT-qPCR和Western blot分析，以及细胞增殖和凋亡测定，研究HOTAIR对ESCs向成肌纤维细胞转分化的影响。结果显示，在用TGF-β1处理的IUA组织和ESC中，HOTAIR的表达水平显着升高。HOTAIR的过表达对ESC具有促纤维化作用，而HOTAIR的沉默则发挥抗纤维化作用。最重要的是，在用pcDNA-HOTAIR转染的TGF-β1处理的ESC中，p-Smad2和p-Smad3的蛋白表达水平显着上调，而在用si-HOTAIR构建体转染后，其蛋白表达水平下调。这些数据表明，HOTAIR通过激活TGF-β1/ Smad信号传导途径促进子宫内膜纤维化，提示HOTAIR的抑制可能代表抑制子宫内膜纤维化的有前途的治疗选择。