MicroRNAs and other non-coding RNAs in Colorectal Metastasis

近30年来，编码蛋白质的癌基因和抑癌基因的改变被认为是肿瘤发生的原因。近年来的研究表明，癌症是一种复杂的遗传性疾病，涉及编码基因和非编码基因的结构和表达异常。我们开创了这样一种想法，即称为microRNA（miRNA）的小非编码RNA（ncRNA）和其他称为超保守基因的较大ncRNA参与人类肿瘤发生，特别是结肠直肠癌（CRC）。肿瘤发生的最后步骤，即转移的发病机制，在很大程度上仍然是未知的。本申请的广泛、长期目的是破译ncRNA在CRC转移过程中的作用，并了解将代表基于miRNA的基因治疗的初始靶点的患者组中的ncRNA遗传异常。为了实现这一目标，我们将使用一种四目标策略，将一大批患者分为两组：一组无转移，另一组有转移。首先，基于微阵列实验与全基因组的miRNA和表达序列标签（EST）分析和生物信息学研究，我们将开发一个大的数据库的miRNA，ultraconserved基因和大ncRNA表达和相关的表达EST的数据库。第二，基于我们将使用生物信息学工具发现的负相关性，我们将分析ncRNA和对mestatases重要的蛋白质编码基因的信使RNA之间可能的相互作用。第三，我们将利用结肠癌转移的体外模型，研究一些差异表达的miRNAs和UCG在转移过程中的生物学功能。基于我们的初步数据，我们将包括作为此类研究的最佳候选者的miR-21、miR-192、miR-215和miR-222，以及非编码UCG uc.160、uc.170A和uc.310，以及根据从患者研究中获得的统计标准选择的前三种miRNA和前三种UCG（如果与先前的基因不同）。所述任务的最终结果将揭示分子诊断和预后的新标志物以及药物治疗的新靶点。

For three decades, alterations of protein-coding oncogenes and tumor suppressors genes have been considered as the causes of tumorigenesis. Recent advances proved without doubts that cancer is a complex genetic disease involving structural and expression abnormalities of both coding and non-coding genes. We pioneered the idea that small non-coding RNAs (ncRNAs) named microRNAs (miRNAs) and other larger ncRNAs named ultraconserved genes are involved in human tumorigenesis and particularly in colorectal cancers (CRC). The pathogenetic mechanisms of the final steps of tumorigenesis, the metastases, is still largely unknown. The broad, long-term purposes of this application are to decipher the roles of ncRNAs during the metastatic process of CRC and to understand the ncRNAs genetic abnormalities in the set of patients that will represent the initial target of miRNA-based gene therapy. To achieve this, we will use a four-aimed strategy applied to a large panel of patients divided in two arms: one without and the other having metastases. First, based on microarray experiments with genome-wide miRNA and expressed sequence tags (ESTs) profiling and on bioinformatics studies, we will develop a large database of miRNAs, ultraconserved genes and large ncRNAs expression and correlate this with databases of expressed ESTs. Second, based on the negative correlations that we will found using bioinformatics tools, we will analyze the possible interactions between ncRNAs and messenger RNAs of protein coding genes significant for mestatases. Third, we will investigate the biological functions related to the metastasis process for some verry significantly differentially expressed miRNAs and UCGs by using in vitro models of colon cancer metastases. Based on our preliminary data, we will include as the top candidates for such studies miR-21, miR-192, miR-215, and miR-222, as well as the non-coding UCGs uc.160, uc.170A and uc.310, and the top three miRNAs and top three UCGs selected according to statistical criteria obtained from the patient study (if different from the previous genes). The final results of the described tasks will reveal new markers for molecular diagnosis and prognosis and new targets for drug therapy.