Development of novel therapy against human liposarcoma using siRNAlibrary

真核细胞中RNA干扰（RNAi）的发现是近年来分子和细胞生物学研究的重大突破。RNAi机制在基因调控、基因组防御、染色质结构和动力学等方面发挥生物学功能。今天，RNAi对动物新型疾病模型的发展产生了巨大的影响。此外，小干扰RNA （sirna）作为RNA沉默的触发分子，很可能成为治疗各种疾病的宝贵工具。所有已知的人黏液样和圆细胞脂肪肉瘤（MLS/RCLS）都与染色体易位有关。这些染色体易位导致编码嵌合癌蛋白的基因融合，该嵌合癌蛋白由两个相关基因TLS（也称为FUS）或EWS中的一个提供的N端和由CHOP（也称为GADD153）基因提供的C端组成。在本项目中，我们试图利用慢病毒siRNA文库鉴定与MLS/RCLS肿瘤发生有关的基因。用慢病毒siRNA文库感染人黏液样脂肪肉瘤细胞系，克隆了部分基因。另一方面，microrna （mirna）是一个21个成员的家族。到25个核苷酸的非编码小rna，这些小rna主要起基因调节作用。MicroRNAs （miRNAs）在细胞增殖、凋亡和分化过程中发挥着重要作用。miRNA的异常表达已被描述为几种人类恶性肿瘤和肿瘤抑制功能已归因于这类新的小调控rna。尽管有这些知识，但关于MLS/RCLS中miRNA的信息缺乏。在本项目中，我们尝试使用微阵列技术鉴定TLS-CHOP的下游miRNA。我们发现了一些调节TLS-CHOP的mirna。有趣的是，我们转染了一些受TLS-CHOP调控的miRNA，抑制了MLS/RCLS细胞系的肿瘤生长。这些数据表明，一些miRNA成为MLS/RCS的可药物靶点。

The discovery of RNA interference (RNAi) in eukaryotic cells is a major breakthrough in the recent progress of the molecular and cellular biology. RNAi machineries exert biological functions in gene regulation, genome defense, chromatin architecture and dynamics. Today, RNAi has had an enormous impact on the development of novel disease models in animals. In addition, small interfering RNAs (siRNAs), the trigger molecules for RNA silencing, are likely to become invaluable tools for the treatment of various diseases.All of known human myxoid and round cell liposarcomas (MLS/RCLS) are associated with chromosomal translocations. These chromosomal translocations lead to gene fusions that encode chimeric oncoproteins consisting of an N terminus contributed by one of two related genes, TLS (also known as FUS) or EWS, and a C terminus contributed by the CHOP (also called GADD153) gene.In this project, we tried to identify the genes that implicated with tumorigenesis of MLS/RCLS using Lentiviral siRNA library. We infected human myxoid liposarcoma cell lines with Lentiviral siRNA library and we cloned some genes.On the other hand, MicroRNAs (miRNAs) are a family of 21. to 25-nucleotide, noncoding small RNAs that primarily function as gene regulators. MicroRNAs (miRNAs) have been suggested to play important roles in cell proliferation, apoptosis, and differentiation. And aberrant miRNA expression has been described for several human malignancies and tumour suppressor functions have been ascribed to this new class of small regulatory RNAs. Despite this knowledge, there is a lack of information regarding miRNA in the MLS/RCLS. In this project, we have tried to identify downstream miRNA of TLS-CHOP using microarray. And we identified some miRNAthat regulated TLS-CHOP. Interestingly, we transfected some of miRNA that regulated by TLS-CHOP, suppressed tumor growth MLS/RCLS cell lines. These data suggested that some miRNA become drugable target for MLS/RCS.