肝癌细胞外泌体诱导STAT3磷酸化促进巨噬细胞极化的分子机制研究  

Molecular mechanism of hepatocellular carcinoma cell exosome-induced STAT3 phosphorylation promoting macrophage polarization

作　　者：燕建洲[1] 杨亚飞 张文权[1] YAN Jianzhou;YANG Yafei;ZHANG Wenquan(Department of Hepatobiliary and Pancreatic Surgery,The NO.2 People's Hospital of Lanzhou,Gansu Lanzhou 730046,China)

机构地区：[1]兰州市第二人民医院肝胆胰外科,甘肃兰州730046

出　　处：《现代肿瘤医学》2025年第6期918-923,共6页Journal of Modern Oncology

摘　　要：目的:探讨肝癌细胞外泌体诱导STAT3磷酸化促进巨噬细胞M2极化的分子机制。方法:通过流式细胞术和实时荧光定量检测巨噬细胞M2极化水平。流式细胞术检测M2型巨噬细胞的细胞表面标志物CD206、实时荧光定量PCR检测M2型巨噬细胞标志物IL-10、ARG1、TGFB1和M1型巨噬细胞标志物表达IL-12A、TNF、NOS2的表达水平。在以下条件下检测:巨噬细胞与正常肝细胞或肝癌细胞共培养后;正常肝细胞或肝癌细胞外泌体处理巨噬细胞后;过表达或敲低FGD5-AS1后;STAT3磷酸化抑制剂SC144或FLLL32处理后。结果:相比于正常肝细胞,与肝癌细胞共培养能够诱导巨噬细胞表达M2型巨噬细胞标志物CD206、IL-10、ARG1、TGFB1(P<0.05),而M1型巨噬细胞标志物表达IL-12A、TNF、NOS2的表达水平下降(P<0.05)。相比于正常肝细胞,肝癌细胞的外泌体可以促进巨噬细胞表达CD206、 IL-10、ARG1、TGFB1(P<0.05),并且抑制IL-12A、TNF、NOS2的表达水平(P<0.05)。在巨噬细胞中敲低FGD5-AS1后,巨噬细胞CD206、IL-10、ARG1、TGFB1的表达受到抑制(P<0.05),并且IL-12A、TNF、NOS2的表达受到促进(P<0.05)。在巨噬细胞中过表达FGD5-AS1后,巨噬细胞CD206、IL-10、ARG1、 TGFB1的表达受到促进(P<0.05),并且IL-12A、TNF、NOS2的表达受到抑制(P<0.05)。过表达FGD5-AS1的正常肝细胞的外泌体处理后,巨噬细胞CD206、IL-10、ARG1、TGFB1的表达受到促进(P<0.05),并且IL-12A、TNF、NOS2的表达受到抑制(P<0.05);敲低FGD5-AS1的肝癌细胞外泌体处理后,巨噬细胞CD206、IL-10、ARG1、TGFB1的表达受到抑制(P<0.05),并且IL-12A、TNF、NOS2的表达受到促进(P<0.05)。在巨噬细胞中敲低FGD5-AS1后,巨噬细胞中STAT3的磷酸化水平下降;在巨噬细胞中过表达FGD5-AS1后,巨噬细胞中STAT3的磷酸化水平上升。STAT3磷酸化抑制剂SC144或FLLL32处理的同时用肝癌细胞的外泌体处理后,巨噬细胞中M2型巨噬细胞标志物CD206、IL-10、ARG1、TGFB1和M1�Objective:To investigate the molecular mechanism of hepatocellular carcinoma cell exosome-induced STAT3 phosphorylation promoting macrophage M2 polarization.Methods:The M2 polarization level of macrophages was quantitatively detected by flow cytometry and real-time fluorescence.The cell surface markers CD206 of M2 macrophages were detected by flow cytometry,and the expression levels of IL-12A,TNF and NOS2 of M2 macrophages markers IL-10,ARG1,TGFB1 and M1 macrophages were detected by real-time quantitative PCR.Under the following conditions:Macrophages were co-cultured with normal hepatocytes or hepatocellular carcinoma cells.Macrophages were treated by exosomes of normal hepatocytes or hepatocellular carcinoma cells.After overexpression or knockdown of FGD5-AS1.STAT3 phosphorylation inhibitor SC144 or FLLL32 treatment.Results:Compared with normal hepatocytes,co-culture with hepatocellular carcinoma cells could induce macrophages to express macrophage markers CD206,IL-10,ARG1 and TGFB1 of M2 type(P<0.05),while the expression levels of IL-12A,TNF and NOS2 of M1 type macrophage markers were decreased(P<0.05).Compared with normal hepatocytes,exosomes of hepatocellular carcinoma cells can promote the expression of CD206,IL-10,ARG1 and TGFB1 in macrophages(P<0.05),and inhibit the expression levels of IL-12A,TNF and NOS2(P<0.05).After FGD5-AS1 was knocked down in macrophages,the expressions of CD206,IL-10,ARG1 and TGFB1 were inhibited(P<0.05),and the expressions of IL-12A,TNF and NOS2 were promoted(P<0.05).After overexpression of FGD5-AS1 in macrophages,the expressions of CD206,IL-10,ARG1 and TGFB1 in macrophages were promoted(P<0.05),and the expressions of IL-12A,TNF and NOS2 were inhibited(P<0.05).After exosome treatment of normal hepatocytes overexpressing FGD5-AS1,the expressions of CD206,IL-10,ARG1 and TGFB1 in macrophages were promoted(P<0.05),and the expressions of IL-12A,TNF and NOS2 were inhibited(P<0.05).The expression of CD206,IL-10,ARG1 and TGFB1 in macrophages was inhibited(P<0.05),and the expression of IL-

关 键 词：肝癌 外泌体 STAT3 磷酸化 巨噬细胞 极化 FGD5-AS1 

分 类 号：R735.7[医药卫生—肿瘤]