Quantitative Proteomics Analysis Reveals Novel Insights into Mechanisms of Action of Long Noncoding RNA Hox Transcript Antisense Intergenic RNA (HOTAIR) in HeLa Cells
Quantitative Proteomics Analysis Reveals Novel Insights into Mechanisms of Action of Long Noncoding RNA Hox Transcript Antisense Intergenic RNA (HOTAIR) in HeLa Cells
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定量蛋白质组学分析揭示了长非编码 RNA Hox 转录反义基因间 RNA (HOTAIR) 在 HeLa 细胞中作用机制的新见解
DOI：
10.1074/mcp.m114.043984
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发表时间：
2015-06-01
影响因子：
7
通讯作者：
Ge, Feng
中科院分区：
文献类型：
--
作者：
Zheng, Peng;Xiong, Qian;Ge, Feng
长链非编码RNA（longnoncodingRNA，IncRNA）是近年来出现的一类重要的基因调控因子，它调控着肿瘤的发生、转移等多种生物学过程。HOTAIR（Hox转录反义基因间RNA）是一种在大多数人类癌症中过表达的lncRNA，已被证明是一种致癌lncRNA。在这里，我们探讨了HOTAIR在HeLa细胞中的作用，并寻找HOTAIR调控的蛋白质。为了从系统的角度理解HOTAIR的作用机制，我们采用定量蛋白质组学策略系统地鉴定HOTAIR的潜在靶点。HOTAIR抑制后，170个蛋白表达显著失调，这意味着它们可能是HOTAIR的潜在靶点。在系统水平上对这些数据的分析揭示了涉及不同细胞成分的蛋白质的重大变化，包括细胞骨架和呼吸链。对细胞骨架中的关键蛋白波形蛋白（vimentin，Vim）的进一步功能研究表明，HOTAIR至少部分通过调节Vim的表达来影响HeLa细胞的迁移和侵袭。HOTAIR的抑制导致线粒体功能障碍和超微结构改变，表明HOTAIR在维持癌细胞线粒体功能中的新作用。我们的研究结果为HOTAIR在癌细胞中的功能机制提供了新的见解。我们期望本研究中所使用的方法将成为IncRNA功能研究的一个组成部分。
Long noncoding RNAs (IncRNAs), 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 IncRNAs.