乳腺肿瘤来源TrxR1促进巨噬细胞免疫抑制性  

Tumor-derived TrxR1 promotes macrophage immunosuppression in breast cancer

作　　者：孙思雨 张松 王岩岩 李煊赫 姜芳倩 姚廷敬 SUN Siyu;ZHANG Song;WANG Yanyan;LI Xuanhe;JIANG Fangqian;YAO Tingjing(Department of Tumor Surgery,the First Affiliated Hospital of Bengbu Medical University,Bengbu 233004,China)

机构地区：[1]蚌埠医科大学第一附属医院肿瘤外科,安徽蚌埠233004

出　　处：《沈阳医学院学报》2025年第2期168-173,共6页Journal of Shenyang Medical College

基　　金：2022年度蚌埠医学院科技项目(No.2022byzd052);2024年度安徽省高等学校科研计划项目(No.2024AH040190)。

摘　　要：目的:探讨TrxR1在乳腺癌中重编程肿瘤相关巨噬细胞的作用及其机制,为乳腺癌的临床治疗提供新的观点和理论依据。方法:利用TISIDB数据库分析TXNRD1 (编码TrxR1)与肿瘤免疫相关性。收集小鼠乳腺癌4T1细胞条件培养基,与骨髓来源巨噬细胞(BMDM)作用48 h,检测巨噬细胞免疫抑制相关因子的表达。通过ELISA试剂盒检测肿瘤细胞TrxR1分泌情况,Westernblot法检测敲低肿瘤细胞TXNRD1的效率。利用荧光定量PCR和流式细胞术进一步检测肿瘤细胞TrxR1的缺乏对巨噬细胞中免疫抑制性因子表达的影响。利用JASPAR数据库分析寻找潜在的调节因子,进一步验证其通路相关蛋白表达和激活情况。结果:数据库表明TXNRD1表达与多种肿瘤生存风险指数成正比,并且TXNRD1与乳腺癌的肿瘤免疫相关性最高。进一步分析发现,随着TXNRD1表达的升高,M1型巨噬细胞和NK细胞浸润减少,而M2型巨噬细胞浸润增多。荧光定量PCR和流式细胞术分析结果显示肿瘤条件培养基可以促进巨噬细胞的免疫抑制性,而敲低肿瘤细胞TXNRD1后的条件培养基则可以抑制巨噬细胞的免疫抑制性。TrxR1中和抗体可明显逆转肿瘤条件培养基对BMDM免疫抑制性因子的诱导作用。利用JASPAR数据库分析得到STAT3和STAT6这2种潜在转录因子,而Western blot法发现敲低肿瘤细胞TXNRD1可以明显抑制巨噬细胞STAT6信号通路的激活。结论:在复杂的肿瘤微环境中,肿瘤来源的TxrR1可促进巨噬细胞免疫抑制性,其机制可能与巨噬细胞中STAT6通路激活相关。Objective:To investigate the role and mechanism of TrxR1 in reprogramming tumor-associated macrophage in breast cancer,providing novel insights and theoretical foundations for clinical breast cancer treatment.Methods:TISIDB database was used to analyze the relationship between TXNRD1(encoding TrxR1)and tumor immunity.Mouse breast cancer 4T1 cells conditioned medium was collected and co-cultured with bone marrow-derived macrophage(BMDM)cells for 48 h to detect the expression of macrophage immunosuppression-related factors.TrxR1 secretion by tumor cells was measured using ELISA kits.TXNRD1 knockdown efficiency was verified via Western blot.Fluorescence quantitative PCR(qPCR)and flow cytometry were used to detect the expression levels of macrophage immunosuppressive factors after TXNRD1 knockdown in tumor cells.JASPAR database was used to analyze the potential regulatory factors,and Western blot was used to verify the expression of pathway-related proteins.Results:Database analysis found that TXNRD1 expression positively correlated with survival risk indices across multiple cancers,with the strongest association observed in breast cancer.Further analysis found that elevated TXNRD1 expression correlated with reduced infiltration of M1 macrophages and natural killer(NK)cells,but increased M2 macrophage infiltration.qPCR and flow cytometry demonstrated that tumor-conditioned medium enhanced macrophage immunosuppression,whereas medium from TXNRD1-knockdown tumor cells suppressed this effect.And TrxR1-neutralizing antibodies could also reversed this effect.JASPAR database analysis identified STAT3 and STAT6 as potential transcriptional regulators,and Western blot confirmed that TXNRD1-knockdown tumor cells conditioned medium inhibited STAT6 pathway activation in macrophages.Conclusion:In the tumor microenvironment,breast tumor-derived TrxR1 promotes macrophage immunosuppression,potentially through activation of the STAT6 signaling pathway.

关 键 词：乳腺癌 TrxR1 肿瘤相关巨噬细胞 微环境 免疫抑制性 

分 类 号：R392.11[医药卫生—免疫学]