miR-373靶向调控JAK2/STAT6信号通路抑制肿瘤相关巨噬细胞M2型极化对直肠癌细胞的影响  

miR-373 inhibits M2 polarization of tumor associated macrophages and affects rectal cancer cells by regulating JAK2/STAT6 signal pathway

作　　者：李鸷[1] 吴迪[1] 谢兴明 田飞[1] 刘洁 LI Zhi;WU Di;XIE Xingming;TIAN Fei;LIU Jie(Department of Gastrointestinal Surgery,the First People’s Hospital of Zunyi,Zunyi 563000,China)

机构地区：[1]遵义市第一人民医院胃肠外科,贵州遵义563000

出　　处：《细胞与分子免疫学杂志》2025年第3期211-220,共10页Chinese Journal of Cellular and Molecular Immunology

基　　金：遵义市科技计划课题[遵市科合HZ字(2019)159号]。

摘　　要：目的 探究miR-373和Janus激酶2/信号转导及转录激活因子6(JAK2/STAT6)信号通路对直肠癌中肿瘤相关巨噬细胞(TAM)向M2型极化的影响。方法 将THP-1细胞诱导为M0/M1/M2型巨噬细胞,M0型巨噬细胞与Caco-2细胞共培养获得TAM,流式细胞术检测CD86、 CD206表达,实时定量PCR、 Western blot法检测miR-373、诱导型一氧化氮合酶(iNOS)、 Toll样受体4(TLR-4)、白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)、精氨酸酶1(Arg1)、几丁质酶样3 (Ym1)、抵抗素样α (Fizz1)、 IL-10 mRNA和蛋白水平。TAM进行细胞转染,分为过表达miR-373组(miR-373-TAM)及其对照组(miR-NC-TAM)、过表达miR-373和JAK2组(miR-373联合JAK2-TAM)及其对照组(miR-373联合NC-TAM),随后与Caco-2细胞共培养。流式细胞术检测TAM中CD206表达;实时定量PCR、 Western blot法检测TAM中miR-373、 Arg1、 Ym1、 Fizz1、 IL-10、 JAK2、 STAT6 mRNA和蛋白水平;CCK-8实验、集落形成实验、 Transwell^(TM)实验检测Caco-2细胞增殖、侵袭和迁移能力。30只裸鼠随机分为Caco-2细胞组、 Caco-2细胞联合miR-NC-TAM组、 Caco-2细胞联合miR-373-TAM组,每组各10只。各组大鼠分别皮下注射单纯Caco-2细胞,Caco-2细胞联合TAM,Caco-2细胞联合过表达miR-373的TAM。细胞接种4周后,免疫荧光染色检测肿瘤组织F4/80^(+)CD206^(+)细胞水平;实时定量PCR、 Western blot法检测肿瘤组织miR-373、 JAK2、 STAT6、 Arg1、 Ym1、 Fizz1、 IL-10 mRNA和蛋白水平。结果 TAM趋向于向M2型极化。过表达miR-373后,TAM中miR-373水平升高,Arg1、 Ym1、 Fizz1、 IL-10、 JAK2、 STAT6 mRNA和蛋白水平降低,Caco-2细胞增殖、侵袭、迁移能力降低;过表达JAK2能够部分逆转过表达miR-373对TAM向M2型极化以及对Caco-2细胞增殖、侵袭、迁移能力的影响。TAM能够促进肿瘤生长;过表达miR-373能够抑制肿瘤生长并抑制TAM向M2型极化。结论 miR-373能够抑制直肠癌中TAM向M2型极化,miR-373可能通过调控JAK2/STAT6通�Objective To explore the effects of miR-373 and Janus kinase 2/signal transducer and activator of transcription 6(JAK2/STAT6)signaling pathways on the M2 polarization of tumor associated macrophages(TAM)in rectal cancer.Methods THP-1 cells were induced into M0/M1/M2 macrophages,M0 macrophages were cocultured with Caco-2 cells to obtain TAM,Flow cytometry was used to detect the expression of CD86 and CD206,Real-time quantitative qPCR and Western blot were used to detect miR-373,inducible nitric oxide synthase(iNOS),toll-like receptor 4(TLR-4),interleukin 1β(IL-1β),tumor necrosis factorα(TNF-α),arginase 1(Arg1),chitinase 3-like 1(Ym1),resistin likeα(Fizz1),IL-10 mRNA and protein levels.TAM were transfected and divided into overexpressing miR-373 group(miR-373-TAM)and control group(miR-NC-TAM),overexpressing miR-373^(+)JAK2-TAM group(miR-373 combined with JAK2-TAM)and control group (miR-373 combined with NC-TAM), and then cocultured with Caco-2 cells. Flow cytometry was used to detect the expression of CD206 in TAM;Real-time quantitative PCR and Western blot were used to detect miR-373, Arg1, Ym1, Fizz1, IL-10, JAK2, STAT6 mRNA and protein levels in TAM;CCK-8 assay, colony formation assay, and Transwell assay were used to detect the proliferation, migration, and invasion ability of Caco-2 cells. Thirty nude mice were randomly divided into Caco-2 cells group, Caco-2 cells combined with miR-NC-TAM group, and Caco-2 cells combined with miR-373-TAM group, with 10 mice in each group. Rats in each group were subcutaneously injected with pure Caco-2 cells, Caco-2 cells combined with TAM, and Caco-2 cells combined with TAM overexpressing miR-373. After 4 weeks of cell inoculation, immunofluorescence staining was used to detect F4/80 ^(+) CD206 ^(+) cells level in tumor tissue;Real-time quantitative PCR and Western blot were used to detect miR-373, JAK2, STAT6, Arg1, Ym1, Fizz1, IL-10 mRNA and protein levels in tumor tissues. Results TAM tended to M2 polarization. After overexpression of miR-373, miR-373 level in TAM wa

关 键 词：直肠癌 miR-373 JAK2/STAT6信号通路 肿瘤相关巨噬细胞 M2型极化 

分 类 号：R392[医药卫生—免疫学] R735.37[医药卫生—基础医学] R714.146