M2型巨噬细胞抑制CD8^(+)T细胞促进食管癌细胞恶性生物行为  

M2 macrophages inhibit CD8^(+)T cells and facilitate the malignant biological behavior of esophageal cancer cells

作　　者：王魏楠 陈素芳 段余钡 井玉莹 黄田平 陈凯 杨凯歌 罗成华 周文虎 胡建明 WANG Weinan;CHEN Sufang;DUAN Yubei;JING Yuying;HUANG Tianping;CHEN Kai;YANG Kaige;LUO Chenghua;ZHOU Wenhu;HU Jianming(Department of Pathology,School of Medicine,Shihezi University/First Affiliated Hospital of Shihezi University,Shihezi 832002;Department of Pharmacy,Xiangya College of Pharmacy,Central South University,Changsha 410013,China)

机构地区：[1]石河子大学医学院病理学系/石河子大学第一附属医院病理科,新疆石河子832002 [2]中南大学湘雅药学院药剂学系,湖南长沙410013

出　　处：《细胞与分子免疫学杂志》2024年第10期887-893,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(82460597);新疆生产建设兵团重点领域科技攻关项目(2023AB058);新疆生产建设兵团科技研究指导项目(2022ZD003,2023ZD028);石河子大学基础研究计划项目(MSPY202407);石河子大学第一附属医院临床基础研究项目(LJ202202)。

摘　　要：目的探讨M2型巨噬细胞通过抑制CD8^(+)T细胞的抗肿瘤能力参与食管癌的恶性生物学行为的作用。方法利用佛波酯(PMA)联合白细胞介素4(IL-4)/IL-13将人单核细胞白血病细胞(THP-1)诱导为M2型巨噬细胞并用实时定量PCR检测相关炎性因子。运用磁珠分选法分选健康志愿者外周血CD8^(+)T细胞,用流式细胞术验证分选纯度。构建CD8^(+)T细胞与食管鳞癌细胞的非接触性共培养体系(CD8^(+)T细胞),和构建M2型巨噬细胞、CD8^(+)T细胞与食管鳞癌细胞的非接触性共培养体系(M2型巨噬细胞联合CD8^(+)T细胞)。平板克隆实验和CCK-8细胞毒性实验检测各组肿瘤细胞增殖能力。Transwell^(TM)实验检测各组肿瘤细胞侵袭和迁移能力。流式细胞术检测并分析各组肿瘤细胞的凋亡情况。运用GraphPadPrism9.5软件进行统计学分析和制图。结果诱导后巨噬细胞IL-10和精氨酸酶1(Arg1)表达上调,IL-12和肿瘤坏死因子α(TNF-α)表达下降,具有M2表型巨噬细胞特征。磁珠分选法成功的分选出CD8^(+)T细胞,其阳性率>90%。食管鳞癌细胞与CD8^(+)T细胞非接触性共培养组的增殖、侵袭、迁移和抗凋亡能力显著低于单独癌细胞组;而食管鳞癌细胞与M2型巨噬细胞条件培养基预处理CD8^(+)T细胞共培养后,其增殖、侵袭、迁移和抗凋亡能力与食管鳞癌和CD8^(+)T细胞共培养组相比显著增强。结论M2型巨噬细胞通过抑制CD8^(+)T细胞的抗肿瘤能力促进食管鳞癌细胞的增殖、侵袭、迁移和抗凋亡。Objective To explore the impact of M2 macrophages on the malignant biological behavior of esophageal cancer by inhibiting the anti-tumor ability of CD8^(+)T cells.Methods Using phorbol myristate acetate(PMA)combined with interleukin 4(IL-4)/IL-13,we induced human monocytic leukemia cells(THP-1)to become M2 macrophages and the detected related inflammatory factors by real-time quantitative PCR.We used magnetic bead sorting to isolate CD8^(+)T cells from healthy volunteers’peripheral blood,and verified the purity of the sorted cells by flow cytometry.We established a non-contact co-culture system between CD8^(+)T cells and esophageal squamous carcinoma cells(CD8^(+)T cell),and established a non-contact co-culture system between M2 macrophages,CD8^(+)T cells,and esophageal squamous carcinoma cells(M2 macrophages combined with CD8^(+)T cell).The plate clone formation assay and CCK-8 cell toxicity assay were used to detect the proliferation ability of tumor cells in each group.The Transwell^(TM) assay was used to detect the invasive and migratory abilities of tumor cells in each group.Flow cytometry was used to detect and analyze the apoptosis of tumor cells in each group.GraphPadPrism9.5 software was used for statistical analysis and graphing.Results After induction,the expression 10期王魏楠,等.M2型巨噬细胞抑制CD8^(+)T细胞促进食管癌细胞恶性生物行为细胞与分子免疫学杂志(Chin J Cell Mol Immunol)2024,40(10)of IL-10 and arginase 1(Arg1)in macrophages was upregulated,while the expression of IL-12 and tumor necrosis factor-alpha(TNF-α)was downregulated,showing the characteristics of M2 macrophages.Peripheral blood CD8^(+)T cells were successfully selected by magnetic bead sorting,with a positive rate of over 90%.The proliferation,invasion,migration and anti-apoptosis ability of esophageal squamous carcinoma cells co-cultured with CD8^(+)T cells in the non-contact manner were significantly lower than those of the single cancer cell group;while the proliferation,invasion,migration and t

关 键 词：M2型巨噬细胞 CD8^(+)T细胞 食管鳞癌 肿瘤微环境(TME) 非接触性共培养 

分 类 号：R735.1[医药卫生—肿瘤] R364.5[医药卫生—临床医学] R392.12