外泌体circ_0001445在胶质瘤内皮细胞血管生成中的调控机制研究

The study of the regulatory mechanism of exosome circ_0001445 in glioma endothelial cell angiogenesis

目的 探索外泌体circ_0001445在胶质瘤内皮细胞血管生成中的调控机制。方法 通过RT-qPCR检测circ_0001445、miR-127-5p在胶质瘤组织中的表达水平。质粒转染T98G胶质瘤细胞以过表达circ_0001445,提取外泌体并鉴定。将外泌体与人脑微血管内皮细胞(HBMECs)培养后,分别通过CCK-8、Transwell小室实验检测过表达T98G细胞外泌体circ_0001445对HBMECs的增殖、迁移、侵袭能力的影响。通过生物信息学技术和双荧光素酶报告基因实验分析circ_0001445和miR-127-5p之间的靶向关系。结果 circ_0001445在胶质瘤组织中显著下调(P<0.05),而miR-127-5p显著上调(P<0.05)。外泌体呈圆形的双层膜囊泡结构,且外泌体标记物CD63和Alix表达均呈阳性。过表达T98G细胞中的circ_0001145可显著抑制HBMECs的增殖、迁移和侵袭能力(P<0.05)。靶向性结果显示,miR-127-5p是circ_0001445的靶标。结论 外泌体circ_0001445可能通过靶向调控miR-127-5p,从而抑制HBMECs的增殖、迁移和侵袭能力。

Objective To explore the regulatory mechanism of exosome circ_0001445 in glioma endothelial cell angiogenesis. Methods The expression levels of circ_0001445 and miR-127-5p in glioma tissues were detected by RT-qPCR. Plasmids were transfected with T98G glioma cells to overexpress circ_0001445, exosomes were extracted and identified. After the exosomes were cultured with human brain microvascular endothelial cells(HBMECs), the effects of overexpression of T98G cell exosome circ_0001445 on the proliferation, migration and invasion of HBMECs were detected by CCK-8 and Transwell assays, respectively. The targeting relationship between circ_0001445 and miR-127-5p was analyzed by bioinformatics techniques and dual-luciferase reporter gene experiments. Results circ_0001445 was significantly down-regulated in glioma tissues(P<0.05), while miR-127-5p was significantly up-regulated(P<0.05). The exosomes were round double-membrane vesicles structure, and the exosome markers CD63 and Alix were both positive. Overexpression of circ_0001145 in T98G cells significantly inhibited the proliferation, migration and invasion ability of HBMECs(P<0.05). The targeting results showed that miR-127-5p was the target of circ_0001445. Conclusion Exosome circ_0001445 may inhibit the proliferation, migration and invasion ability of HBMECs by targeting regulating miR-127-5p.