乙型肝炎性肝癌性别差异的假基因活化与长链非编码RNA调控网络及功能研究

项目名称： 乙型肝炎性肝癌性别差异的假基因活化与长链非编码RNA调控网络及功能研究

项目编号： No.81330037

项目类型： 重点项目

立项/批准年度： 2014

项目学科： 医药、卫生

项目作者： 孙树汉

作者单位： 中国人民解放军第二军医大学

项目金额： 290万元

中文摘要： 慢性乙型肝炎相关肝细胞癌（HCC）的发生与预后存在显著性别差异，男女发病约3—5：1。流行病调查和基础实验均证实，雌性激素对肝癌具有保护作用，雄性激素促进男性的肝癌发生。但针对性激素各个环节的干预手段对其防治却没有任何效果。因此我们需要从一个新的角度对这一问题进行重新审视。近年来越来越多的研究表明，曾经一度被人们忽视的长链非编码RNA(long noncoding RNA，lncRNA)、假基因等分子在生理以及各种疾病状态下都有重要的调控功能。本项目计划首先利用新一代测序技术筛选肝癌组织中特征性表达的lncRNA、假基因，对关键性lncRNA、假基因的结构、相互结合的蛋白、DNA分子、参与调控的信号转导通路以及假基因活化、lncRNA作为肝癌早期诊断、疾病进展预测标记分子的可能性等方面进行深入的探讨。为该疾病的早期诊断、预后判断提供一组新的生物标记物，并为其治疗提供新的潜在药物作用靶点。

中文关键词： C09_肝和肝内胆管肿瘤;长链非编码RNA;性别;表观遗传学;新一代测序

英文摘要： The major risk factors for HCC are chronic liver diseases with cirrhosis that include hepatitis B. Epidemiological reports indicate that regardless of etiologies, the incidence of HCC is higher in males than in females with the male: female ratio usually averaging between 3-5:1. In view of this remarkable gender disparity, various in vitro as well as in vivo studies have been initiated from time to time to explore the importance of sex hormones in HCC. Though the studies began as early as in 1980s, the precise role of sex hormones and the significance of their receptors in HCC still remain poorly understood and perhaps contribute to current controversies about the potential use of hormonal therapy in HCC. We need a new perspective to explore this issue. The human transcriptome comprises not only large numbers of protein-coding messenger RNAs (mRNAs) but also a large set of non-protein coding transcripts that have structural, regulatory or unknown functions. A surprisingly wide array of cellular functions has also been associated with long non-coding RNAs (lncRNAs, tentatively defined as non-coding RNAs more than 200 nt in length) and pseudogenes. A handful of studies have implicated lncRNAs and pseudogenes in a variety of disease states, and altered lncRNA and pseudogene levels can result in aberrant expression of gene products that may contribute to cancer biology. However, the overall pathophysiological contributions of lncRNAs and pseudogenes to HCC remain largely unknown. .In this study, we plan to identify non-overlapping signatures of a small number of lncRNAs and pseudogenes that are up-regulated or down-regulated in HBV-related HCC compared with paired peritumoral tissues. Previous studies indicate that lncRNAs are key regulators of chromatin state, yet the nature and sites of RNA-chromatin interaction are mostly unknown. Here we introduce chromatin isolation by RNA purification, where tiling oligonucleotides retrieve with key lncRNAs bound protein and DNA sequences, which are enumerated by mass spectrometry identifying and deep sequencing. The studies presented here were also designed to test whether monitoring the relative expression of key lncRNA and pseudogenes and related pathways in clinic HCC samples will help to predict the prognosis of HCC. Together, our objective here is to highlight the important roles of lncRNA and pseudogene in the development and progress of HCC and test whether it is possible to apply lncRNAs and pseudogenes in HCC therapy and prediction of HCC prognosis.

英文关键词： liver and intrahepatic bile du;long noncoding RNA;gender;epigenetics;next-generation Sequencing