结直肠癌发生相关lncRNAs的筛选、验证及其血清学诊断价值研究

Long noncoding RNAs (lncRNAs), a novel class of noncoding RNAs, were recently found to be dysregulated in various diseases and have critical regulatory roles in wide array of cellular functions. The altered expression of lncRNAs can result in aberrant gene expression products which may contribute to cancer biology. However, the overall pathophysiological contributions of lncRNAs to tumorigenesis of colorectal cancer (CRC) have not been fully elucidated and it is also of paramount important to develop novel molecular markers for CRC diagnosis based on differentiate lncRNAs profiles, which could help patients prevent delay of effective treatment. To define associations between lncRNAs and CRC, we sough to identify the specific lncRNAs expression profiles based on the thesis of "normal mucosa-adenoma-carcinoma" to evaluate the function and diagnostic value. In our previous studies, high-throughput lncRNAs chip analysis was performed in tissue samples of CRC patients, adenoma patients and healthy controls. Thirty-nine lncRNAs were found differently expressed. In further studies, we will verify the 39-lncRNAs profile in a large cohort and establish the specific lncRNAs expression profile. The vitro assays including cell proliferation, colony formation, cell apoptosis, cell cycle and the vivo nude mice experiments combined with lncRNA-mRNA coexpression analysis will be used to demonstrate the role of lncRNAs in the tumorigenesis of CRC. In addition, based on the important advances with the introduction of exosomes, we will further assess the diagnostic values of differential lncRNAs by RT-qPCR in serum exosomes, which may show us medically values on early detection of CRC.

长链非编码RNAs（lncRNAs）是新发现的具有基因表达调控作用的一类分子，其异常表达可导致肿瘤的发生，而它们与结直肠癌发生的关系以及是否可作为分子标志物用于肿瘤诊断迄今尚未阐明。本课题组前期利用高通量lncRNAs芯片初筛了39个结直肠癌、结直肠腺瘤及癌旁正常组织差异表达lncRNAs。为此，本研究拟扩大临床样本进行验证，建立结直肠癌发生相关lncRNAs差异表达谱；然后通过体外细胞实验、体内裸鼠模型及lncRNA-mRNA共表达分析，探讨差异lncRNAs在结直肠癌发生中的作用；最后以血清exosomes为检测载体，运用RT-qPCR技术对结直肠癌、结直肠腺瘤以及健康对照者血清进行大样本、多层次验证，评价血清exosomes中差异lncRNAs对结直肠癌的临床诊断价值及转化医学意义。本研究不仅有助于进一步明确lncRNAs在结直肠癌发生中的作用，也将为结直肠癌的实验室诊断提供依据。

结直肠癌是发于末端消化道的恶性肿瘤，是目前最常见的恶性肿瘤之一,发病率直肠癌高于各类型肠道肿瘤。长链非编码RNAs（lncRNAs）是新发现的具有基因表达调控作用的一类分子，其异常表达可导致肿瘤的发生，而它们与结直肠癌发生的关系以及是否可作为分子标志物用于肿瘤诊断迄今尚未阐明。本课题组前期利用高通量lncRNAs芯片初筛了多个结直肠癌、结直肠腺瘤及癌旁正常组织差异表达lncRNAs，如NR_026817, NR_029373, NR_034119, SPINT1-AS1, CRNDE-h, HOTAIR, MALAT1等。在此基础上，本研究通过扩大临床样本进行验证，建立结直肠癌发生相关lncRNAs差异表达谱；然后通过体外细胞功能实验及敲低/获得性共表达分析，探讨差异多个lncRNAs在结直肠癌增殖，转移，化疗耐药等发生中的作用，分析其在结直肠癌诊断及预后预测中的价值；最后以血清外泌体为检测载体，运用RT-qPCR技术对外泌体CRNDE-h在结直肠癌、结直肠腺瘤以及健康对照者血清进行稳定性及表达水平分析，验证了血清外泌体lncRNA CRNDE-h对结直肠癌的临床诊断及预后价值。本项目研究不仅能为结直肠癌的早期筛查和疗效预测开辟新的途径，而且对结直肠癌治疗方案的选择具有重要的指导意义。