ceRNA MALAT1竞争性结合miR-200s调控角膜新生血管及新生淋巴管形成的机制研究

The research on the pathogenesis and treatment of corneal neovascularization and cornea lymphangiogenesis has been a hot and difficult problem in ophthalmology. Some studies show that vascular endothelial growth factor(VEGF) plays an important role in the physiological and pathological process of angiogenesis and lymphangion. Through the analysis of bioinformatics, we found that there are some binding sites of miR-200 family molecules in the gene sequence of VEGF. In addition, there are some interaction between ceRNA MALAT1 and miR-200s. Therefore, we plan to investigate the effect of on the expression of miR-200s and VEGF by regulating the expression of MALAT1, using Prox1-GFP / Flt1-DsRed (PGFD) transgenic fluorescent mice, respectively, from the level of animal and in vitro cell level, using luciferase reporter gene system, quantitative PCR technology, protein immune technology, cell experiments and pathological detection, etc. By interfering MALAT1 expression, we will investigate the effect of miR-200s and VEGF expression, the proliferation, apoptosis, migration and new blood vessel and lymphangiogenesis formation. The molecular mechanisms were explored and combined with the clinical data analysis, for the clinical prevention and treatment of corneal neovascularization and lymphangiogenesis to explore new therapeutic targets and directions for exploratory research, and provide experimental data support.

角膜新生血管和新生淋巴管的发病机制和治疗方法一直都是眼科的热点和难点。许多研究表明血管内皮生长因子（VEGF）在血管及淋巴管生成的生理及病理过程中发挥重要调控作用。申请人通过多个数据库进行生物信息学分析发现miR-200家族分子与VEGF mRNA均存在结合位点，而ceRNA MALAT1与miR-200s存在相互作用。由此，我们计划通过干预MALAT1的表达，利用Prox1-GFP/Flt1-DsRed (PGFD)转基因荧光小鼠，从动物水平和细胞水平借助荧光素酶报告基因系统、定量PCR技术、蛋白免疫技术、细胞实验及病理检测等手段，分别探讨其对miR-200s、VEGF水平的影响，对内皮细胞增殖、凋亡、迁移及新生血管和淋巴管形成的影响；从中发掘相关分子机制，结合临床样本检测数据分析，为角膜新生血管和新生淋巴管的临床预防及治疗发掘新的治疗靶点及方向进行探索性研究，并提供实验数据支持。

角膜新生血管和新生淋巴管的发病机制和治疗方法一直都是眼科的热点和难点。研究表明血管内皮生长因子（VEGF）在血管及淋巴管生成的生理及病理过程中发挥重要调控作用。我们通过生物信息学分析发现miR-200家族分子与VEGF mRNA均存在结合位点，而ceRNA MALAT1与miR-200s存在相互作用。我们通过改变MALAT1的表达，利用转基因荧光小鼠，发现其变化显著影响角膜新生血管和新生淋巴管，确定在小鼠角膜新生血管和新生淋巴管中MALAT1与miR-200s、VEGF表达水平的相关性。进一步验证miR-200s对VEGF的转录调控作用，以及MALAT1表达下调对miR-200s和VEGF相关基因及蛋白表达水平、角膜新生血管新生淋巴管形成的影响。lncRNA MALAT1作为竞争性内源性RNA，可能通过miRNA-200s来发挥作用影响角膜新生血管和新生淋巴管。