烟草致癌物NNK诱发长链非编码RNA基因突变在肺癌发生中的作用

项目名称： 烟草致癌物NNK诱发长链非编码RNA基因突变在肺癌发生中的作用

项目编号： No.81472937

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 杨巧媛

作者单位： 广州医科大学

项目金额： 75万元

中文摘要： 肺癌是对人类生命威胁最大的恶性肿瘤之一，目前普遍认为环境致癌因素致相关蛋白编码基因突变是肺癌启动的分子基础，然而对环境致癌物诱发长链非编码RNA基因突变及其功能的研究尚未见报道。本研究通过检测烟草致癌物NNK诱导肺上皮细胞恶性转变后lncRNA表达谱的变化，采用多种生物信息学方法，综合分析筛选NNK相关的lncRNA基因作为捕获区域，通过目标序列捕获测序技术广泛寻找lncRNA基因突变位点，根据突变位点的重要性评分进行突变位点的筛选，并在肺癌人群中进行验证；通过构建不同基因型细胞和对不同基因型肺癌病例临床资料的分析，从细胞和人群水平定量分析NNK诱发的lncRNA基因突变对lncRNA表达调控和肺癌发生发展的影响，为环境致癌的分子机制和肺癌的病因学研究提供新的思路和实验依据。

中文关键词： 肺癌；长链非编码RNA；基因突变；化学致癌；表观遗传调控

英文摘要： Lung cancer is one of the most common cancers in the world and the leading cause of cancer deaths.Development of lung cancer depends on both genetic and environmental factors.The mutations of environmental response genes are the basis of lung cancer development. Non-coding RNA(ncRNA) is an important kind of new environmental response genes discovered in recent years. Although many classes of functional ncRNAs have been recently described(including microRNAs, snRNAs and circular RNAs), the function of long noncoding RNA (lncRNA) have remained elusive in the field of chemical carcinogenesis.In the current study, we identified specific lncRNA related with the induction of tabacco specific carcinogen NNK[4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone] by using lncRNA microarray technology and bioinformatical analysis. Then,these specific lncRNA genes were sequenced by sequence capture array to discover the mutations within these genes. Some important mutations were selected to verify in lung cancer patients. Then,the candidated mutations were introduced into the cell model by recombinant DNA technique and site-directed mutagenesis.Subsequently, we observed the effect of lncRNA gene mutation on the carcinogenesis of lung cancer induced by NNK in vitro. The epidemiological study was also carried out to explore the relationship between lncRNA gene mutation and lung cancer risk in vivo. We hope that the current study will provide novel insights into the fields of environmental carcinogenesis and cancer pathogenesis.

英文关键词： lung cancer;long non-coding RNA;gene mutation;chemical carcinogenesis;epigenetic regulation