Department of Radiation Oncology, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, China.
Core Facility Center, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, China.
Cancer Med. 2023 Jun;12(11):12608-12621. doi: 10.1002/cam4.5941. Epub 2023 Apr 25.
Nasopharyngeal carcinoma (NPC) is a type of head and neck cancer with high incidence in China. The molecular mechanisms of vasculogenic mimicry (VM) and angiogenesis are not fully elucidated in NPC. More specially, it has seldomly been reported that Epstein-Barr virus-encoded miRNA can regulate VM and angiogenesis in NPC. The aim of this study was to investigate the function and molecular mechanism of a targeting exosome system (iRGD-exo-antagomiR) against VM and angiogenesis in NPC, and to provide new approaches for improving the comprehensive treatment of NPC.
Exosomes were isolated by differential ultracentrifugation. Dynamic light scattering, transmission electron microscopy and western blotting were performed to characterize the exosomes. The 3D-Culture assay, tube formation assay, chicken chorioallantoic membrane assay, Matrigel plug assay, mouse xenograft tumor modeling and immunohistochemical staining were applied to evaluate the anti-VM and anti-angiogenic effects of the targeting exosome system in vitro and in vivo. Western blot was performed to detect the changes of downstream regulated networks following interference and recovery of the target gene.
In vitro or in vivo treatment with iRGD-tagged exosome containing antagomiR-BART1-5p specifically suppressed VM and angiogenesis in NPC. EBV-miR-BART1-5p promoted VM and angiogenesis in vitro and in vivo by regulating VEGF, PI3K, Akt, mTOR and HIF1-α in a Spry2-dependent manner.
Our findings demonstrated that targeting exosomes enveloped EBV-miR-BART1-5p-antagomiRs in a Spry2-dependent manner for NPC therapy through both anti-VM and anti-angiogenesis in vitro and in vivo.
鼻咽癌（NPC）是一种在中国发病率较高的头颈部癌症。血管生成拟态（VM）和血管生成的分子机制在 NPC 中尚未完全阐明。更特别的是，很少有报道称 Epstein-Barr 病毒编码的 miRNA 可以调节 NPC 中的 VM 和血管生成。本研究旨在探讨针对 NPC 中 VM 和血管生成的靶向外泌体系统（iRGD-exo-antagomiR）的功能和分子机制，并为提高 NPC 的综合治疗提供新的途径。
通过差速超速离心法分离外泌体。动态光散射、透射电子显微镜和 Western blot 用于表征外泌体。3D 培养实验、管形成实验、鸡胚尿囊膜实验、Matrigel plugs 实验、小鼠异种移植肿瘤建模和免疫组织化学染色用于评估靶向外泌体系统在体外和体内的抗 VM 和抗血管生成作用。Western blot 用于检测干扰和恢复靶基因后下游调控网络的变化。
体外或体内用 iRGD 标记的含有 antagomiR-BART1-5p 的外泌体治疗特异性抑制 NPC 中的 VM 和血管生成。EBV-miR-BART1-5p 通过调节 VEGF、PI3K、Akt、mTOR 和 HIF1-α，以 Spry2 依赖的方式在体外和体内促进 VM 和血管生成。
我们的研究结果表明，针对外泌体包裹的 EBV-miR-BART1-5p-antagomiRs 的靶向治疗通过体外和体内的抗 VM 和抗血管生成作用，为 NPC 治疗提供了一种新的方法。
