蒽环类抗肿瘤药物的免疫激活作用及其机制  

Immune activation of anthracycline drugs and mechanism

作　　者：黄欧[1] 姜敏[2] Huang Ou;Jiang Min(Department of General Surgery,Comprehensive Breast Health Center,Ruijin Hospital,Shanghai Jiaotong University School of Medicine,Shanghai 200025,China;Department of Medical Oncology,the First Affiliated Hospital of Soochow University,Suzhou 215006,China)

机构地区：[1]上海交通大学医学院附属瑞金医院外科乳腺疾病诊治中心,上海200025 [2]苏州大学附属第一医院肿瘤科,苏州215006

出　　处：《中华实验外科杂志》2023年第4期651-654,共4页Chinese Journal of Experimental Surgery

摘　　要：目的探讨蒽环类药物除了损伤细胞DNA杀伤肿瘤细胞外,是否具有诱导肿瘤细胞免疫原性死亡激活抗肿瘤免疫反应。方法采用THP1-Dual单核细胞中干扰素刺激基因(ISG)的报告系统,检测蒽环类药物伊达比星、柔红霉素和阿霉素对ISG的影响,并采用荧光定量聚合酶链反应(PCR)的方法检测蒽环类对干扰素(IFNB、IFNA4)和趋化因子IP-10的基因表达影响,以及检测伊达比星对多种肿瘤细胞IFNB的表达影响。采用蛋白免疫印迹的方法,检测蒽环类化合物对干扰素上游TBK1/IRF3信号通路的影响。最后,采用ATM抑制剂和TBK1抑制剂和STING敲除的THP1-Dual报告细胞,分析伊达比星是否通过DNA损伤感受通路刺激IFNB的生成。采用单因素方差分析。结果蒽环类药物具有促进ISG的表达及干扰素基因(IFNB)生成的作用,其中伊达比星的效果最强,在1μmol/L下作用24 h后对ISG激活约5倍(P<0.01),在1μmol/L下作用6 h后对IFNB、IP-10和IFNA4基因激活分别达21.4、3827.2、4.3倍(P<0.01)。在多种肿瘤细胞株中,伊达比星能促进KG-1和JIMT-1中IFNB基因的表达,在3μmol/L作用6 h后分别达28.0、81.3倍(P<0.01)。蒽环类化合物促进TBK1/IRF3磷酸化信号通路促进干扰素的生成,并通过使用ATM抑制剂(AZ31、KU55933)、TBK1抑制剂(MRT67307、BX795)和STING敲除的细胞模型,发现伊达比星促进干扰素通路是依赖于ATM、TBK1和STING分子,提示蒽环类药物通过诱导DNA损伤感受通路促进ATM的活化及STING/TBK1/IRF3通路,进而促进干扰素的生成。结论蒽环类药物特别是伊达比星具有诱导免疫原性细胞死亡的作用,它们通过诱导DNA损伤感受通路间接激活抗肿瘤免疫反应。Objective To investigate whether anthracycline drugs,in addition to damaging the DNA of cancer cells to kill them,also induce immunogenic cell death and activate anti-tumor immune responses.Methods A reporter system for interferon-stimulated genes(ISGs)in THP1-Dual monocytes was used to detect the effects of the anthracycline drugs(idarubicin,doxorubicin and aclarubicin)on ISGs.The quantitative fluorescent polymerase chain reaction(PCR)was used to detect the effects of anthracyclines on the gene expression of interferons(IFNB and IFNA4)and chemokine IP-10,and to examine the effects of idarubicin on IFNB expression in various cancer cells.Additionally,immunoblotting was used to investigate the effects of anthracyclines on the TBK1/IRF3 signaling upstream pathway of interferons.Finally,the study analyzed whether idarubicin stimulated IFNB production via the DNA damage sensing pathway using ATM and TBK1 inhibitors and THP1-Dual KO-STING cells.One way ANOVA was used to test the effects of anthracycline drugs on gene expressions.Results Anthracycline drugs promoted the expression of ISGs and the production of interferon genes(IFNB),with idarubicin having the strongest effect.After 24 h of treatment with 1μmol/L idarubicin,ISG activation increased by approximately 5-fold,and after 6 h of treatment at 1μmol/L,activation of IFNB,IP-10 and IFNA4 gene increased by 21.4-fold,3827.2-fold,and 4.3-fold,respectively.In multiple cancer cell lines,idarubicin promoted IFNB gene expression in KG-1 and JIMT-1 cells,with fold increases of 28.0 and 81.3(P<0.01),respectively.After 6 h of treatment at 3μmol/L,anthracyclines promoted the phosphorylation of the TBK1/IRF3 signaling pathway,and using inhibitors of ATM(AZ31,KU55933),TBK1(MRT67307,BX795),and STING knockdown cell models,the study found that idarubicin promoted the interferon pathway dependently on ATM,TBK1,and STING molecules.These findings suggest that anthracycline drugs promote the activation of ATM and STING/TBK1/IRF3 pathways by inducing the DNA damage sensing pathway,

关 键 词：蒽环类药物 抗肿瘤免疫反应 干扰素 

分 类 号：R730.5[医药卫生—肿瘤]