Exosomes synergized with PIONs@E6 enhance their immunity against hepatocellular carcinoma via promoting M1 macrophages polarization
Exosomes synergized with PIONs@E6 enhance their immunity against hepatocellular carcinoma via promoting M1 macrophages polarization
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DOI：
10.1016/j.intimp.2021.107960
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发表时间：
2021-07-18
影响因子：
5.6
通讯作者：
Wen, Jian
中科院分区：
文献类型：
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作者：
Chen, Hanren;Jiang, Shulian;Wen, Jian
背景：肝细胞癌（HCC）由于促进M2型巨噬细胞极化而促肿瘤发生，缺乏抗肿瘤免疫力，术后容易复发。近年来的研究表明，外泌体与HCC的发生、发展密切相关。来自 HCC 的抗原外泌体能够极化为替代激活的巨噬细胞 M2，但不会刺激 M1 巨噬细胞极化。氧化铁纳米粒子 (ION) 已被证明能够促进 M1 巨噬细胞极化。本研究旨在探索外泌体作为载体与负载二氢卟酚 e6 的聚乙二醇化离子 (PIONs@E6) 协同作用，通过促进 M1 巨噬细胞极化来增强其对 HCC 的免疫力。材料和方法：合成PIONs@E6，然后分别通过化学物理分析、透射电子显微镜（TEM）进行表征。通过TEM对含有PIONs的外泌体进行表征后，然后通过Western Blotting测定外泌体表面特异性分子CD9和CD63。分别通过酶联免疫吸附测定 (ELISA) 和流式细胞术分析体外和体内 M1 巨噬细胞极化标志物。使用 Spectra Max 荧光酶标仪分析巨噬细胞中的细胞内活性氧 (ROS)。通过监测体内异种移植小鼠模型中的肿瘤生长来评估含有 PIONs 的外泌体对 HCC 的抑制作用。结果：PIONs@E6表现出良好的水溶性，核心直径约为10 nm，水合物直径约为37 nm。外泌体特异性标志物CD9和CD63的表达保持在高水平。含有 PION 的外泌体可以剂量依赖性地促进体外和体内 M1 巨噬细胞极化。值得注意的是，含有 PIONs 的外泌体可以显着提高巨噬细胞中的 ROS 水平，并显着抑制 HCC 异种移植小鼠的肿瘤生长。结论：外泌体作为载体可以与PIONs@E6协同作用，通过促进M1巨噬细胞极化来增强对HCC的免疫力。
Background: Hepatocellular carcinoma (HCC) is easy to relapse after resection for its lack of anti-tumor immunity due to pro-tumorigenesis by promoting M2 type macrophage polarization. Recent studies have shown that exosomes are closely related to the occurrence and development of HCC. Antigenic exosomes from HCC are able to polarize into alternatively activated macrophages M2, but do not stimulate M1 macrophages polarization. Iron oxide nanoparticles (IONs) have been demonstrated to be able to promote M1 macrophages polarization. This research was to explore exosomes as vehicles to synergize with pegylated IONs loaded with chlorin e6 (PIONs@E6) to enhance their immunity against HCC via promoting M1 macrophages polarization. Materials and Methods: PIONs@E6 was synthesized and then characterized by chemico-physical analysis, transmission electron microscope (TEM), respectively. After characterization of PIONs-contained exosomes by TEM, and then the exosomal surface specific molecules CD9 and CD63 were determined by Western Blotting assay. Markers of M1 macrophage polarization in vitro and in vivo were analyzed by enzyme linked immunosorbent assay (ELISA) and flow cytometry, respectively. Intracellular reactive oxygen species (ROS) in macrophages were analyzed using a Spectra Max fluorescence microplate reader. Inhibitory effect of PIONs-contained exosomes on HCC was evaluated by monitoring tumor growth in an in vivo xenograft mice model. Results: PIONs@E6 showed good water solubility with a core diameter around 10 nm and a hydrate diameter around 37 nm. The expression of exosome specific markers CD9 and CD63 was kept at a high level. PIONscontained exosomes can dose-dependently promote M1 macrophages polarization in vitro and in vivo. Of note, PIONs-contained exosomes could initiate a significantly higher level of ROS in macrophages and remarkably inhibit the tumor growth in mice bearing HCC xenograft. Conclusion: Exosomes as vehicles could be synergized with PIONs@E6 to enhance their immunity against HCC via promoting M1 macrophages polarization.