环境致癌中基因组水平非编码序列表遗传修饰图谱的构建及特征功能分析

The molecular mechanism of lung cancer induced by exogenous environmental carcinogen is the research focus in lung cancer etiology and environmental medicine. As a kind of active environmental response gene, many non-coding RNAs(ncRNA) are regulated by environmental carcinogens and play an important role in environmental carcinogenesis. However, there is not enough attention on the molecular mechanism  of ncRNA disregulation induced by environmental carcinogens. In our present study, we use a NNK-induced malignant transformated normal human bronchial epithelial cells as a model and establish the environmental specific ncRNA database. We plan to examine the methylation level of ncRNA gene by MeDIP sequencing and ncRNA promoters methylation array. Then, the methylated and demethylated cell models will be constructed in our study to quantitative analyse the the impact of methylation modification on ncRNA expression and lung cancer carcinogenesis. We will verify our results of in vitro experiments in lung cancer population. A comprehensive map of genetic and epigenetic regulatory database on ncRNA will be bulit in combination with our previous research. It will provide important resources and experimental basement to further clarify the role of ncRNA in environmental carcinogenesis.

非编码RNA（ncRNA）转录于基因组非编码区，是一类活跃的环境应答基因，从ncRNA基因水平探索ncRNA上游调控机制是申请人长期致力的研究方向。本课题组前期主要关注ncRNA基因突变的遗传调控机制，研究结果提示甲基化等表遗传修饰可能是调控环境致癌中ncRNA异常表达的另一重要途径。本研究以代表性环境致癌物NNK染毒建立恶性转化细胞模型，联合全基因组DNA甲基化测序、ncRNA测序及启动子甲基化芯片等高通量技术，筛选环境特异性非编码序列，在基因组水平建立环境特异性非编码序列甲基化和乙酰化等表遗传修饰图谱；预测功能性位点，体外构建甲基化和去甲基化细胞模型，从细胞和人群水平定量分析表遗传修饰对非编码序列转录水平和对肺癌发生发展的影响；结合前期研究，分析环境致癌中基因组编码区遗传突变和表遗传修饰全貌，寻找并明确重点调控位点的功能，为环境致癌分子机制研究和暴露效应靶点的寻找开辟新的领域。

非编码RNA（ncRNA）是一类活跃的环境应答基因，甲基化修饰是其重要的上游调控模式。本项目以代表性环境致癌物NNK染毒人正常肺支气管上皮细胞，建立恶性转化细胞模型；联合全基因组DNA甲基化测序、ncRNA测序及启动子甲基化芯片等高通量技术，在基因组水平建立了环境特异性非编码序列甲基化表遗传修饰图谱；筛选出环境特异性非编码RNA lncRNA AC007255.8，分析了甲基化修饰水平对lncRNA AC007255.8转录表达水平和对肺癌发生发展的影响，并在肺癌人群中进行了初步验证，从细胞和人群水平定量分析表遗传修饰对非编码序列转录水平和对肺癌发生发展的影响，为环境致癌分子机制研究和暴露效应靶点的寻找开辟新的领域。项目项目经费使用合理，按期完成项目计划书的研究目标和研究任务，研究结果已发表中英文论文7篇，其中SCI收录英文5篇，中文核心期刊2篇，培养硕士研究生4人，申请发明专利1项，作为第二完成人获得省部级科技奖励2项，目前尚有部分研究结果在投稿中。