长链非编码RNA HOTAIR与YBX1（Y-box protein-1）蛋白相互作用调控肿瘤细胞增殖的分子机制研究

Numerous long non-coding RNAs (lncRNAs) were found to play significant roles in cancer progression. LncRNA HOTAIR (HOX antisense intergenic RNA) is dys-regulated in various kinds of cancer cells, and HOTAIR can exert its function in cancer cells by directly interacting with proteins. In our previous study, we have systematically identified HOTAIR--interacting proteins by using interatomic analysis, and found that YBX1 was hubs in the HOTAIR interacting-protein networks. The cellular function of YBX1 depends on its location and distribution in thenucleus or in the cytoplasm, and our preliminarystudies found that HOTAIR can repress the translocation of YBX1 from cytoplasm to nucleus. However, it is still unclear that how HOTAIR influence the nuclear translocation of YBX1, and how HOTAIR promote proliferationof cancer cells by interacting with YBX1?Super-resolution imaging and affectfunctional studies will be carried out to fully understand the mechanism of how HOTAIR affect the proliferation of cancer cells by regulating YBX1.We expect that new therapy targets for cancer will be found and the strategy used in this study will become an integral part of functional studies of lncRNAs-protein interaction.

长链非编码RNA(lncRNA)是一种具有复杂生物学功能的，转录本长度超过200个核苷酸的RNA分子，HOTAIR (HOX antisense intergenic RNA)是一种在多种肿瘤中异常表达的lncRNA，可通过与多种蛋白的相互作用来调控肿瘤细胞的增殖。在前期研究中，我们首次发现HOTAIR可与YBX1（Y-box protein-1）蛋白相互作用并调控其入核，YBX1蛋白是一个具有多种重要生物学功能的转录因子，参与调控肿瘤细胞的增殖。本研究拟综合采用分子生物学、功能蛋白质组学和分子影像技术，阐明HOTAIR调控YBX1入核的分子机制；揭示HOTAIR如何通过与YBX1相互作用来调控肿瘤细胞的增殖及其具体的分子机制；进一步通过对多种不同的肿瘤细胞的分析，明确该现象和机制在肿瘤细胞中是否具有普遍性。本项目的实施将提出HOTAIR调控肿瘤细胞增殖的新的分子机制。

本研究的研究目标是合采用分子生物学、功能蛋白质组学和分子影像技术，阐明HOTAIR调控YBX1入核的分子机制；揭示HOTAIR如何通过与YBX1相互作用来调控肿瘤细胞的增殖及其具体的分子机制；进一步通过对多种不同的肿瘤细胞的分析，明确该现象和机制在肿瘤细胞中是否具有普遍性。目前以上目标已经全部完成。本项目利用蛋白质组学的方法，系统鉴定HOTAIR在hela细胞中的相互作用蛋白； 构建首个以细菌光敏色素荧光蛋白为基础的三分子荧光系统(iRFP-TriFC)并用于细胞内验证lncRNA-蛋白的相互作用；为lncRNA-蛋白研究提供新方法； 抑制HOTAIR表达对YBX1的磷酸化及入核具有调控作用；HOTAIR 通过pAKT、pERK 调控 YBX1入核；发现了HOTAIR通过调控YBX1的磷酸化及入核，上调细胞增殖正调控因子PCK2, PDGFRB的表达，进而促进细胞的增殖。