HCG11靶向结合miR-543调节血肿瘤屏障通透性  

HCG11 regulates the blood-tumor barrier permeability by targeting miR-543

作　　者：郭吉喆 赫倩茹 刘丽波 阮雪蕾 马珺 薛一雪 GUO Ji-zhe;HE Qian-ru;LIU Li-bo;RUAN Xue-lei;MA Jun;XUE Yi-xue(Department of Neurobiology,College of Basie Medieine,China Medical University,Shenyang 110122,China)

机构地区：[1]中国医科大学基础医学院神经生物学教研室,沈阳110122

出　　处：《解剖科学进展》2018年第6期588-592,共5页Progress of Anatomical Sciences

基　　金：国家自然科学基金(81573010);辽宁省科学技术计划项目(No.2017225020);沈阳市科技计划项目(No.17-231-1-46)

摘　　要：目的研究HCG11和miR-543对血肿瘤屏障通透性的作用及可能机制。方法应用Real-time PCR检测人脑微血管内皮细胞(endothelial cells, ECs)和人胶质瘤微血管内皮细胞(glioma endothelial cells, GECs)中HCG11和miR-543的表达以及二者的结合作用。应用Lipo3000将HCG11表达沉默和/或miR-543过表达质粒载体分别转染GECs,验证转染效率后,通过测量跨内皮细胞阻抗值(transendothelial electric resistance, TEER)和辣根过氧化物酶(horseradish peroxidase, HRP)渗透量,检测血肿瘤屏障通透性。应用Real-time PCR和Western blot检测紧密连接相关蛋白ZO-1、Occludin和Claudin-5的表达。荧光素梅报告基因系统分析miR-543与HCG11的结合作用。结果 HCG11在GECs中表达显著增加,miR-543在GECs中表达显著降低;HCG11表达沉默、miR-543过表达均显著降低TEER值,增加HRP渗透量,降低ZO-1、Occludin和Claudin-5的蛋白表达水平。MiR-543与HCG11存在靶向结合作用。HCG11表达沉默显著增加miR-543的表达,增加血肿瘤屏障通透性。结论 HCG11通过靶向结合miR-543调节血肿瘤屏障通透性。Objective To investigate the effect and possible mechanism of HCG11 and miR-543 on blood tumor barrier permeability. Methods Real-time PCR was used to detect the expression of HCG11 and miR-543 in human brain microvascular endothelial cells(ECs) and human glioma endothelial cells(GECs) and the interaction between HCG11 and miR-543. HCG11 silencing plasmid vectors and/or miR-543 agomir were transfected into GECs. After the transfection the efficiency was tested, transendothelial electric resistance(TEER) and horseradish peroxidase(HRP) flux assays were used to test the permeability of BTB. The mRNA and protein expression levels of tight junction related proteins ZO-1, Occludin and Claudin-5 were evaluated by Real-time PCR and Western blot respectively. Dual-luciferase reporter assay was performed to explore potential interactions between HCG11 and miR-543. Results The expression level of HCG11 was significantly increased, while miR-543 was decreased in GECs. Knockdown of HCG11 or overexpression of miR-543 presented a decrease in TEER and an increase in HRP flux, and decreased the expression levels of ZO-1, occludin and claudin-5. MiR-543 functionally targeted HCG11, and knockdown of HCG11 significantly increased the expression of miR-543 and permeability of blood tumor barrier. Conclusion HCG11 regulated the blood-tumor barrier permeability by targeting miR-543.

关 键 词：HCG11 miR-543 血肿瘤屏障 紧密连接 

分 类 号：R730.2[医药卫生—肿瘤]