肿瘤研究方法-Approaches-and-Techniques.ppt

肿瘤的实验动物和细胞研究技术 The animal model and in vitro research of tumor 甲基化检测方法 Hpa II +Msp I 酶切法 Hpa II → -CCGG- ↗ Msp I ↘-CCmGG- 通过电泳分析酶切片段即可知甲基化状态 Suggested topics be focused on in the discussion course What will you add to the list of the approaches and techniques that I have presented, if you have, please give some details. What is your Ph.D. research proposal? What kinds of methods and techniques you intend to use in your research? What are the problems you may encounter and how to solve them? How much do you know about RNAi technique? Will you give us some points. The connection of microRNA with tumor formation MiRNAs are negative regulators of gene expression MicroRNAs (miRNAs) are a group of non-coding RNA, to date, more than 200(239) miRNAs have been described in humans The levels of some miRNAs are altered in cancer continued The most convincing example is B cell lymphoma: There are extra copies in chromosome 13q-31-32 in B cell lymphoma, this region contains a gene called c13 (f25), which turns out to encode the precursors of 7 miRNAs These 7 miRNAs (called mir-17-92 polycistron) are overexpressed in B cell lymphoma Enforced expression of mir-17-92 ploycistron acts with c-myc expression can increase in the frequency of the lymphoma in mice ( from about 30% to 100%) and decrease the latency of the disease ( from 3-6 m to 51 days) continued MicroRNAs expression profile may used to classify poorly differentiated tumors ,which highlight the potential of miRNA profiling in cancer diagnosis 四、蛋白组学及生物芯片 蛋白组学（proteomics) Y-轴等电电泳或固态pH 2-D gel electrophoresis 梯度电泳 X-轴SDS银染或荧光染色，找出差异点，挖胶 ↓ 质谱分析，微量氨基酸测序 ↓ BLAST确定是已知还是未知的基因，克隆该基因 生物芯片（biochip) 是指采用光导原位合成或微量点样等方法将大量核酸片段（寡核苷酸、cDNA、基因组DNA、RNA）或多肽分子甚至细胞等生物样品有序地固定于支持物（玻片、硅片、聚丙烯酰胺凝胶、尼龙膜等载体）的表面，组成密集的二维分子排列，然后与已标记的待测生物样品中靶分子杂交，通过特定