缺氧肝细胞癌细胞来源外泌体miR-432-5p通过SOCS3/STAT3促进M2型巨噬细胞极化

Hypoxia-induced exosome miR-432-5p promotes M2 macrophage polarization through SOCS3/STAT3 signal pathway in hepatocellular carcinoma

目的探讨缺氧微环境下肝细胞癌(简称肝癌)细胞释放的外泌体对巨噬细胞极化的影响及其机制。方法通过免疫荧光染色、透射电镜、Western blotting明确缺氧对肝癌细胞外泌体释放的影响;通过流式细胞术、免疫荧光染色和实时定量PCR检测缺氧肝癌细胞来源外泌体对M2型巨噬细胞极化、免疫微环境的影响;通过免疫组织化学染色、免疫荧光染色、荧光素酶报告基因、Western blotting检测缺氧肝癌细胞来源外泌体miR-432-5p促进M2型巨噬细胞极化的机制。结果缺氧可促进肝癌细胞分泌外泌体,促进M2型巨噬细胞极化,其发挥作用的机制是外泌体中的miR-432-5p靶向SOCS3,调控SOCS3/STAT3信号通路实现的。结论缺氧肝癌细胞来源外泌体miR-432-5p介导的M2型巨噬细胞极化是通过SOCS3/STAT3信号通路实现的,这一信号通路为肝癌的靶向治疗提供了新方向。

Objective To investigate the effect and mechanism of hepatocellular carcinoma exocrine release on macrophage polarization in a hypoxic microenvironment.Methods The effect of hypoxia on the exosome release of hepatocellular carcinoma cells was determined using immunofluorescence staining,transmission electron microscope,and Western blotting.Flow cytometry,immunofluorescence staining,and real-time quantitative polymerase chain reaction were used to detect the effects of hypoxia hepatoma cell-derived exocrine on M2 macrophage polarization and an immune microenvironment.Immunohistochemical staining,immunofluorescence staining,luciferase reporter gene,and Western blotting were used to detect the mechanism by which hypoxia hepatoma cell exocrine miR-432-5p promotes M2 macrophage polarization.Results Hypoxia can promote M2 macrophage polarization and hepatocelluar carcinoma cells for exosome secretion.The mechanism for this is through the miR-432-5p in exosomes,which target SOCS3 to regulate the SOCS3/STAT3 signal pathway.Conclusion M2 macrophage polarization mediated by miR-432-5p exocrine from hypoxic hepatocellular carcinoma cells is realized through SOCS3/STAT3 signal pathway,which offers a new direction for targeted therapy of hepatocellular carcinoma.