中科院水生所关于长链非编码RNA的研究取得重要进展

　　

   长链非编码RNA(Long non-coding RNA, lncRNA)是长度大于 200 个核苷酸的非编码 RNA。研究表明lncRNA作为一种重要的基因调控模式参了多种生物过程例如表观遗传调控、细胞周期调控、肿瘤发生和转移等。通过研究lncRNA的生物学功能及其在疾病中的调控机制，能够更加全面深入地理解疾病发生的分子机理，并有望发现新的疾病诊断标志物以及治疗靶点，是目前生命科学领域的研究热点。

   中国科学院水生生物研究所葛峰课题组采用基于生物质谱分析的体内标记定量蛋白质组学技术，系统、深入研究了一种重要的肿瘤相关LncRNA即HOTAIR的蛋白调控网络；将SILAC（stable isotope labeling by amino acids in cell culture）方法应用于lncRNA的功能研究中，构建了HOTAIR的蛋白调控网络（图1），生物信息学分析发现HOTAIR参与调控多种重要生物学过程，尤其是细胞骨架和线粒体相关的生理活动。

   深入的功能研究结果表明，HOTAIR可以通过上调一种关键的细胞骨架蛋白—vimentin (VIM)的表达来调控肿瘤细胞的迁移和侵袭.抑制HOTAIR的表达还会导致线粒体的功能紊乱以及超微结构的变化，提示HOTAIR的正常表达在维持癌细胞线粒体功能方面也起着重要作用。通过整合定量蛋白质组学和功能研究的结果，研究人员提出了HOTAIR调控肿瘤细胞迁移和侵袭的分子机制模型（图2）。

   研究结果表明，SILAC技术是系统研究和鉴定非编码RNA的调控蛋白网络和功能靶标的高效且可靠的方法，可以从动态和整体的角度阐明细胞或生物体非编码RNA的蛋白调控网络和分子作用机制，有望成为非编码RNA功能研究的不可或缺的工具.该研究成果已经在线发表在国际蛋白质组学专业杂志Molecular & Cellular Proteomics上。文章链接: http://www.mcponline.org/content/early/2015/03/11/mcp.M114.043984.long

 

 
图1.基于SILAC的定量蛋白质组学技术研究一种长链非编码RNA的蛋白调控网络的实验路线及结果.

 
图2.通过整合定量蛋白质组学和细胞实验以及动物实验的结果，提出了一种长链非编码RNA-HOTAIR调控肿瘤细胞迁移和侵袭的分子机制模型.

原文摘要：

Quantitative  Proteomics  Analysis  Reveals  Novel  Insights  into Mechanisms of Action of Long Non-coding RNA HOTAIR in HeLa Cells
Long non-coding RNAs (lncRNAs), which have emerged in recent years as a new and crucial layer of gene regulators, regulate various biological processes such as carcinogenesis and metastasis. HOTAIR (Hox transcript antisense intergenic RNA), a lncRNA overexpressed in most human cancers, has been shown to be an oncogenic lncRNA. Here, we explored the role of HOTAIR in HeLa cells and searched for proteins regulated by HOTAIR. To understand the mechanism of action of HOTAIR from a systems perspective, we employed a quantitative proteomic strategy to systematically identify potential targets of HOTAIR. The expression of 170 proteins was significantly dys-regulated after inhibition of HOTAIR, implying that they could be potential targets of HOTAIR. Analysis of this data at the systems level revealed major changes in proteins involved in diverse cellular components, including the cytoskeleton and the respiratory chain. Further functional studies on vimentin (VIM), a key protein involved in the cytoskeleton, revealed that HOTAIR exerts its effects on migration and invasion of HeLa cells, at least in part, through the regulation of VIM expression. Inhibition of HOTAIR leads to mitochondrial dysfunction and ultrastructural alterations, suggesting a novel role of HOTAIR in maintaining mitochondrial function in cancer cells. Our results provide novel insights into the mechanisms underlying the function of HOTAIR in cancer cells. We expect that the methods used in this study will become an integral part of functional studies of lncRNAs.