外泌体与PIONs@E6协同作用，通过促进M1巨噬细胞极化增强其对肝细胞癌的免疫作用。
Exosomes synergized with PIONs@E6 enhance their immunity against hepatocellular carcinoma via promoting M1 macrophages polarization.
作者信息
Chen Hanren, Jiang Shulian, Zhang Peng, Ren Zhongyu, Wen Jian
机构信息
Guilin Medical University, Guilin, Guangxi, People's Republic of China.
Nanjing Second Hospital, Nanjing, Jiangsu, People's Republic of China.
出版信息
Int Immunopharmacol. 2021 Oct;99:107960. doi: 10.1016/j.intimp.2021.107960. Epub 2021 Jul 18.
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. PIONs-contained 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.
背景
肝细胞癌（HCC）因通过促进M2型巨噬细胞极化而具有促肿瘤发生作用，缺乏抗肿瘤免疫，故切除术后易复发。近期研究表明，外泌体与HCC的发生发展密切相关。HCC来源的抗原性外泌体能极化成为交替活化的M2巨噬细胞，但不刺激M1巨噬细胞极化。氧化铁纳米颗粒（IONs）已被证明能够促进M1巨噬细胞极化。本研究旨在探索以外泌体为载体，与负载二氢卟吩e6的聚乙二醇化IONs（PIONs@E6）协同作用，通过促进M1巨噬细胞极化增强其对HCC的免疫作用。
材料与方法
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PIONs的外泌体进行表征后，再用蛋白质免疫印迹法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巨噬细胞极化的标志物。使用Spectra Max荧光酶标仪分析巨噬细胞内的活性氧（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PIONs的外泌体在体外和体内均可剂量依赖性地促进M1巨噬细胞极化。值得注意的是，含PIONs的外泌体可在巨噬细胞中引发显著更高水平的ROS，并显著抑制荷HCC异种移植小鼠的肿瘤生长。
结论
外泌体作为载体可与PIONs@E6协同作用，通过促进M1巨噬细胞极化增强其对HCC的免疫作用。
Zotero 插件