食管癌中非编码RNA调控的代谢通路高异常区域识别及其分子互作机制

In recent years, our experimental results showed that thousands of ncRNAs were significantly differentially expressed in esophageal cancer, and played an important role in regulating metabolic pathways of abnormal cancer cells such as glucose or lipid metabolism. However, for so many ncRNAs, which one can directly target key enzymes and regulate metabolic pathways of abnormal cancer cells is still an important scientific problems in doubt. Therefore, the project plan to firstly establish a new direct model of metabolic pathways with ncRNAs as key regulatory nodes. Secondly, the significantly differential ncRNAs, protein-coding genes and metabolites in esophageal cancer are mapped to the above model of metabolic pathways. Through integrative analyses and development of the new algorithm of bioinformatics, the high abnormal regions and subpathways in metabolic pathways can be identified, which will further help to locate key ncRNAs in the metabolic pathways and the key enzymes directly regulated by them. Finally, through applying an integrative and comparative strategy to overexpression and knockdown experiment, the project will finalize the accurate interactions between ncRNA and key metabolic enzymes and the corresponding functions to the experimental confirmation, which will further confirm scientific hypothesis of abnormal molecular mechanism of metabolic pathways synergistically regulated by ncRNAs at the molecular/cell/animal/clinical tissue level.

近年，我们的实验结果表明，数以千计的ncRNA在食管癌中显著差异表达，在癌细胞糖或脂等代谢通路异常中发挥重要调控作用。然而，对于如此众多的ncRNA，究竟谁直接靶向代谢通路关键代谢酶，调控癌细胞代谢通路异常这一重要科学问题尚不清楚。为此，本项目拟首先建立以ncRNA为关键调控节点的有向代谢通路新模型；其次，将食管癌中显著差异表达的ncRNA、蛋白编码基因以及代谢子等信息，投射到上述代谢通路模型中予以系统整合分析，开发生物信息学新算法，精细识别食管癌代谢通路中的高异常区域，以及高异常区域代谢子通路，并进一步确定代谢子通路中的ncRNA及其直接调控的关键代谢酶；最后，通过综合运用基因高表达与敲降表达对比实验策略，在分子/细胞/动物实验/临床组织水平,对前述确定下来的ncRNA调控关键代谢酶表达的精细互作情况及其功能予以实验确认，证明食管癌中ncRNA协同调控代谢通路异常分子机制科学假说。

研究显示，数以千记的miRNA和lncRNA等ncRNA在肿瘤组织细胞中异常表达。这些差异表达的ncRNA，很可能通过调控通路中的关键蛋白编码基因（PCG），参与肿瘤细胞对重要通路的异常调控。代谢通路中的基因能够编码重要的代谢相关蛋白和关键代谢酶，很可能受到ncRNA的调控。本项目根据研究计划，建立了以ncRNA为关键调控节点的通路新模型平台。在该平台下，本项目成功建立了以癌症lncRNA为代表的ncRNA有向代谢通路模型。接下来，通过将癌症显著差异表达的ncRNA、PCG以及代谢相关信息，映射到通路模型中进行系统分析，开发了精细识别癌症通路高异常区域的新方法。通过多种癌症数据的处理和分析，验证了方法的有效性。以食管癌为模型，本项目挖掘了食管癌高异常子通路，并进一步确定了通路中的关键功能的ncRNA，以及ncRNA直接调控的关键基因、蛋白和代谢酶。而且，我们发现了许多功能性的ncRNA被增强子，尤其超级增强子调控。这表明增强子和超级增强子能够通过控制ncRNA的转录水平，继而调控多个下游通路，放大其对癌症的影响力。最后，我们综合利用基因敲降和过表达等实验技术和策略，在分子/细胞/临床组织/动物水平,对生物信息学方法预测的重要ncRNA进行功能的验证，并进一步揭示食管癌中ncRNA调控通路的分子机制。综上所述，本项目的顺利完成，对揭示食管癌等癌症的分子互作和通路机制及探索癌症生物分子网络分析方法具有重要的意义。在本项目的研究期间，我们已经发表成果论文多篇,并构建了支持食管癌等多种癌症的ncRNA调控通路识别软件和分析平台。