MicroRNAs and other non-coding RNAs in Colorectal Metastasis

三十年来，编码蛋白质的癌基因和抑癌基因的改变一直被认为是肿瘤发生的原因。最近的研究证明，癌症是一种复杂的遗传病，涉及编码和非编码基因的结构和表达异常。我们率先提出了这样的想法，即被称为microRNAs(MiRNAs)的小非编码RNA(NcRNAs)和其他被称为超服务基因的较大ncRNAs参与了人类肿瘤的发生，特别是在结直肠癌(CRC)中。肿瘤发生的最后几步--转移--的致病机制在很大程度上仍不清楚。这一应用的广泛、长期的目的是破译ncRNAs在结直肠癌转移过程中的作用，并了解一组患者中ncRNAs的遗传异常，这将是基于miRNA的基因治疗的初始靶点。为了实现这一点，我们将使用四个目标的策略应用于分为两个手臂的一大群患者：一支没有转移，另一支有转移。首先，基于全基因组miRNA和表达序列标签(ESTs)图谱的微阵列实验和生物信息学研究，我们将建立一个关于miRNAs、超服务基因和大型ncRNAs表达的大型数据库，并将其与EST表达数据库相关联。其次，基于我们将利用生物信息学工具发现的负相关性，我们将分析对Mestatase有重要意义的蛋白质编码基因的ncRNAs和信使RNAs之间可能的相互作用。第三，我们将利用结肠癌体外转移模型，研究一些差异表达的miRNAs和UCGs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个miRNAs和前三个UCG(如果与先前的基因不同)，作为此类研究的首选候选。上述任务的最终结果将揭示分子诊断和预后的新标记物，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.