非编码基因与肿瘤分析.ppt

1. microRNA的研究历史 2. microRNA分布和作用机制 3. microRNA与癌症 4. microRNA研究的一般步骤和方法 5 以一篇文章为例介绍一下研究的过程 2001年 命名为microrna The first demonstration of a link between miRNA genes and cancer. The first quantitative real-time PCR developed for miRNA profiling, designed for the amplification of precursor molecules. An elegant study that shows a pathogenetic link between miRNAs and target oncogenes. The first paper to show that the deregulation of a single miRNA gene can lead to cancer. microrna与癌症 The let-7 family negatively regulates Ras The miRNAs that are encoded by the let-7 family were the first group of oncomirs shown to regulate the expression of an oncogene, specifically the Ras genes. RAS原癌基因参与细胞的增殖和分化， 约有15-30%的肿瘤中含有RAS基因的突变 Ras家族有3个成员，即Nras、Kras、Hras分别含有9个、8个和3个let-7的靶位点，在HepG2细胞系中过表达let-7显著降低RAS蛋白的水平，在HeLa细胞系中转染let-7的反义抑制分子，则会提高RAS蛋白的水平 の miR-17–92簇 miR-17–92簇位于13q31，其中包含了7个microRNA: miR-17-5p, miR-17-3p, miR-18, miR-19a, miR-20, miR-19b-1 和miR-92-1 ODonnell 66等发现MYC可以结合到miR-17-92簇所在基因C13orf25的第一个内含子上，并诱导miR-17-92簇的表达 4. microRNA研究的一般步骤和方法 4. microRNA研究的一般步骤和方法 Real-time PCR Real-time PCR Microarray 5 以一篇文章介绍一下研究的过程 Out of a total of eight selected miRNAs, three (miR-155, miR-9 and miR-10b) were found to be markedly upregulated in breast cancer cells when compared with either primary human mammary epithelial cells (HMECs) or with the spontaneously immortalized MCF- 10A cells 5.1 miR-10b? cell migration and invasion in vitro 5.2 miR-10b ? tumour invasion in vivo 5.3 miR-10b ? distant metastasis in vivo 5.4 how miR-10b expression is regulated? 5. 5 What is a direct and functional target of miR-10b? 5.6 miR-10b expression is elevated in metastatic breast tumours 5.1 miR-10b? cell migration and invasion in vitro first performed in vitro loss-of-function analyses by silencing the miRNAs with antisense oligonucleotides suggesting that each of the transfected antisense RNAs achieved a greater than 50% inhibition of the actions of its cogn