长链非编码RNA-VEC1340靶定KLF4在血管内皮细胞损伤中的调控及机制研究

项目名称： 长链非编码RNA-VEC1340靶定KLF4在血管内皮细胞损伤中的调控及机制研究

项目编号： No.81470550

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 赵朝

作者单位： 西安交通大学

项目金额： 69万元

中文摘要： 研究表明长链非编码（lncRNA）在各种生物学过程及疾病中发挥重要的调节作用。lncRNA-VEC1340是我们前期筛选并初步验证在损伤的人脐静脉内皮细胞（HUVEC）中异常表达的lncRNA，并通过生物信息学和文献分析预测Krüppel样因子4（KLF4）是其靶基因。本项目拟对lncRNA-VEC1340在HUVEC损伤中的作用及机制开展进一步研究：（1）通过gain-of-function和loss-of-function干预实验明确VEC1340在HUVEC损伤中的作用；（2）荧光素酶报告基因系统、RNA pull-down、竞争性结合实验验证KLF4 是其靶标；（3）Western blot、免疫共沉淀、激光共聚焦显微镜研究VEC1340对KLF4表达水平、蛋白半衰期、乙酰化及亚细胞定位的影响。通过以上研究将为VEC损伤的治疗提供新的理论基础、实验依据及治疗靶点。

中文关键词： 长链非编码RNA；Krüppel样因子4；动脉粥样硬化；血管内皮细胞

英文摘要： An increasing body of evidence reveals that lncRNA plays an important role in modulating a varity of biological processes and is involved in the pathogenesis of many diseases. lncRNA-VEC1340, preliminarily selected and validated by IncRNA chip in our pre-experiment, was abnormally expressed in the injured HUVEC, and KLF4 was predicted to be the target gene by bioinformatics and literature analysis. This project was to carry out further studies on the role and mechanism of lncRNA-VEC in the injured HUVEC: (1) To define the role of lncRNA-VEC1340 in the damage of HUVEC by using the experiments of gain-of-function and loss-of-function. (2) To verify the predicted tagart KLF4 through Luciferase@ Reporter Gene System, RNA pull-down and competitive binding experiment. (3)To explore the regulation of KLF4 expression levels, half-life and acetylation of protein, sbucelluar location by lncRNA-VEC1340 through the Western blot, co-immunoprecipitation and laser scanning confocal microscope. It will provide new theoretical basis, experimental evidence and therapeutic targets through this project.

英文关键词： lncRNA;Krüppel-like factor 4;atherosclerosis;vascular endothelia cell