长链非编码RNA-CACS2及其启动子区多态性与结直肠癌发生发展的相关性及作用机制研究

项目名称： 长链非编码RNA-CACS2及其启动子区多态性与结直肠癌发生发展的相关性及作用机制研究

项目编号： No.81201581

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 肿瘤学1

项目作者： 陈典克

作者单位： 中山大学

项目金额： 23万元

中文摘要： 长链非编码RNA(LncRNA) CASC2参与肿瘤发生发展，但作用机制不明。申请者前期工作发现LncRNA CASC2在肠癌组织中表达下调，其低表达与启动子区高甲基化相关，且位于启动子区CpG岛的单核苷酸多态性（SNP）亦与肠癌相关。综合分析以上结果，我们提出"CASC2启动子区的SNPs通过影响CpG岛甲基化调控CASC2表达，调节CASC2的重要调控网络，从而与肠癌发生发展密切相关"的科学假设。本项目拟进一步扩大样本验证该假设，在体外细胞实验中，通过构建重组质粒、荧光素酶报告基因、EMSA/ChIP实验获得SNP位点改变CpG岛甲基化状态参与调控CASC2的具体机制；采用基因克隆、siRNA干扰技术获得CASC2不同表达水平的肠癌细胞模型，通过一系列细胞学实验观察其增殖、分化、侵袭、转移等细胞表型改变。最后采用荷瘤裸鼠等动物体内实验确证CASC2在肠癌中的作用，并探讨其潜在治疗效果。

中文关键词： 长链非编码RNA；结直肠癌；UCA1；临床预后；侵袭转移

英文摘要： The long non-coding RNA (LncRNA) CASC2 involved in cancer development, but the effect and mechanism was unknown. In our previous research we have found LncRNA CASC2 down-regulated in colorectal cancer(CRC) tissue, its low expression associated with promoter hypermethylation. And then we perform a screening to investigate the association polymorphisms in the promoter region of the LncRNA CASC2 with the risk of CRC in a hospital based case-control study. The results shown that some single nucleotide polymorphism(SNP)located within the promoter CpG island were associated with susceptibility toCRC.Based on the previous research results, ,we proposed the scientific hypothesis that CASC2 promoter region polymorphism located CpG island, may modify the state of methylation ,involving in the regulation of CASC2 expression levels, and thus affecting important regulatory network participation by CASC2 , which is closely related to colorectal cancer development. In order to verify this hypothesis. The project intends to confirm the interaction among LncRNA CASC2 promoter region polymorphism, CpG island methylation state, expression levels, and their association with CRC development by further expand sampls. In vitro, the construction of recombinant plasmid, Site-Directed Mutagenesis, double luciferase reporter gene assay, e

英文关键词： Long no-coding RNA；colorectal cancer；Urothelial Cancer Associated 1；Clinical prognosis；Invasion and metastasis