circ-PTBP3对血肿瘤屏障通透性的影响  

Effect of circ-PTBP3 on the Permeability of Blood-Tumor Barrier

作　　者：马珺[1] 姚一龙[2] 阮雪蕾 沈书园 邵连奇 薛一雪[1] MA Jun;YAO Yilong;RUAN Xuelei;SHEN Shuyuan;SHAO Lianqi;XUE Yixue(Department of Neurobiology,School of Life Sciences,China Medical University,Shenyang 110122,China;Deparment of Neurosurgery,Shengjing Hosipital,China Medical University,Shenyang 110004,China)

机构地区：[1]中国医科大学生命科学学院神经生物学教研室,沈阳110122 [2]中国医科大学附属盛京医院神经外科,沈阳110004

出　　处：《中国医科大学学报》2019年第12期1057-1062,共6页Journal of China Medical University

基　　金：国家自然科学基金（81602725,81573010）;辽宁省自然科学基金重点项目（20170520441）;辽宁省博士科研启动基金（201601123,20170520047）;沈阳市科技计划（17-231-1-46）

摘　　要：目的探讨环状RNA(circRNAs)circ-PTBP3(hsa_circ_0088072)在胶质瘤微血管内皮细胞(GECs)中的表达,以及circ-PTBP3对血肿瘤屏障(BTB)通透性的调控作用。方法构建血脑屏障与BTB模型,应用qRT-PCR检测正常脑微血管内皮细胞(ECs)与GECs中circ-PTBP3的表达水平。RNase R验证circ-PTBP3对其的抵抗作用。建立circ-PTBP3稳定沉默的GECs,用跨内皮阻抗值(TEER)检测屏障的完整性,通透性测定检测屏障的通透性,Western blotting和免疫荧光实验检测紧密连接相关蛋白ZO-1、occludin与claudin-5的表达与分布。结果circ-PTBP3在GECs中高表达,沉默circ-PTBP3可显著降低BTB的TEER值,增加BTB的通透性,减少紧密连接相关蛋白ZO-1、occludin与claudin-5的表达与分布。结论沉默circ-PTBP3可显著增加BTB的通透性,减少紧密连接相关蛋白的表达。circ-PTBP3可能成为开放BTB的分子靶标,有效提高胶质瘤的治疗效果。Objective To investigate the expression of circ-PTBP3(hsa_circ_0088072)in glioma microvascular endothelial cells(GECs)and the regulation of blood-tumor barrier(BTB)permeability by circ-PTBP3.Methods The blood-brain barrier and BTB models were constructed to estimate the expression levels of circ-PTBP3 in normal brain microvascular endothelial cells(ECs)and GECs by qRT-PCR.RNase R was used to verify the resistance of circ-PTBP3 to degradation.GECs with stably silenced circ-PTBP3 were generated.The integrity and permeability of the BTB were measured by transendothelial electric resistance(TEER)and permeability assays,respectively.The expression and distribution of tight junction-related proteins ZO-1,occludin,and claudin-5 were detected by Western blotting and immunofluorescence assays,respectively.Results circ-PTBP3 was found to be highly expressed in GECs.Silencing circ-PTBP3 significantly reduced the TEER value and increased the permeability of BTB.This was accompanied by a decrease in the expression and distribution of the tight junction-related proteins ZO-1,occludin,and claudin-5.Conclusion Silencing circ-PTBP3 significantly increased BTB permeability,and reduced the expression of tight junction-related proteins.Therefore,circ-PTBP3 may serve as a molecular target for BTB opening,and this would in turn effectively improve the efficacy of therapeutics targeting glioma.

关 键 词：环状RNA 血肿瘤屏障 通透性 胶质瘤 

分 类 号：R34[医药卫生—基础医学]