外泌体科学研究策略与方法.pptx

外泌体研究策略;目录 一、外泌体概述 二、外泌研究兴起 三、外泌体基础研究技术 四、外泌体研究策略基本逻辑 五、外泌体研究实例;外泌体概述;外泌体研究兴起;外泌体中标项目机制研究包括主要非编码RNA种类;;外泌体研究基础技术;外泌体研究策略基本逻辑;外泌体研究基本路线图;外泌体研究整体研究策略;实例一 由生物分子到外泌体;Exosome-Transmitted lncARSR Promotes Sunitinib Resistance in Renal Cancer by Acting as a Competing Endogenous RNA;1、lncARSR Is Highly Expressed in Sunitinib-Resistant RCC Cells;2、lncARSR Is Modulated by AKT/FOXO Axis in Sunitinib-Resistant RCC Cells;3、lncARSR Is Required for Sunitinib Resistance of RCC;4、lncARSR Levels in Plasma and Tumor Tissues Correlate with Sunitinib Response in RCC Patients;5、Packaging of lncARSR into Exosomes Is Mediated by hnRNPA2B1;6、Intercellular Transfer of lncARSR by Exosomes Disseminates Sunitinib Resistance;7、AXL and c-MET Are Responsible for lncARSR-Mediated Sunitinib Resistance;8、lncARSR Functions as a ceRNA for miR-34 and miR-449 to Facilitate AXL and c-MET Expression;21/38;9、Targeting lncARSR Restores Sunitinib Response in RCC;总结： 这篇文章中外泌体不是主角，外泌体只是作为长非编RNA功效作用机制一个主要方面，但总体上给我们展现了外泌体研究一系列主要方法： 1）外泌体功效探讨基础：供体细胞（舒尼替尼抗性肾癌细胞）/受体细胞（舒尼替你敏感性肾癌细胞）。 2）外泌体功效前期预试验：功效生物分子位于外泌体中（图5A）、外泌体电镜检测（图5B）、不一样起源外泌体中功效分子差异表示（图5C）、功效分子被特异包裹入外泌体关键蛋白检测（图5D）。 3）外泌体功效试验：外泌体处理细胞、外泌体进细胞核以及受供体细胞共孵育等。 ;实例二由外泌体到生物分子 ;Endothelial microparticles mediate inflammation-induced vascular calcification;1、TNF-a up-regulates EC BMP-2 expression through activation of NF-kB and stimulates the generation of EMPs;2、Annexin V, calcium, and BMP-2 in EMPs from HUVECs stimulated by TNF-a;3、The effect of EMPs from HUVECs stimulated by TNF-a on VSMC calcification;4、Knockdown of BMP-2 in HUVECs produces nonmineralizing TNF-a-EMPs;5、EMPs from patients with CKD stimulate VSMC calcification and osteogenesis And from senescent HUVECs induce calcification and osteogenic changes of VSMCs;6、Internalization of EMPs by VSMCs;实例三 ;CD90+ liver cancer cells modulate endothelial cell phenotype through the release of exosomes containing H19 lncRNA;1、CD90+ cells show a mesenchymal phenotype and actively release exosomes;2、Exosomes released by