EGFR-TKI通过cGAS-STING信号通路对肺癌小鼠放疗远隔效应的影响  

EGFR-TKI enhances abscopal effect of radiotherapy in mice with lung cancer via cGAS-STING signaling pathway

作　　者：韩有溪 马荣辉[2] 艾秀清[1] 朱相露[1] 王义海 HAN Youxi;MA Ronghui;AI Xiuqing;ZHU Xianglu;WANG Yihai(Department of Breast Radiotherapy,The Affiliated Tumor Hospital of Xinjiang Medical University,Urumqi 830000,China;不详)

机构地区：[1]新疆医科大学附属肿瘤医院乳腺放疗科,乌鲁木齐830000 [2]新疆医科大学附属肿瘤医院呼吸神经内科

出　　处：《山东医药》2024年第31期45-50,共6页Shandong Medical Journal

基　　金：新疆医科大学附属肿瘤医院人才项目(2021KX005);新疆维吾尔自治区自然科学基金资助项目(2021D01C419)。

摘　　要：目的探讨表皮生长因子受体酪氨酸激活抑制剂(EGFR-TKI)通过环鸟苷酸-腺苷酸合成酶-干扰素基因刺激因子(cGAS-STING)信号通路对肺癌小鼠放疗远隔效应的影响。方法取部分对数生长期肺癌细胞(LLC1细胞)随机分为对照组和辐照组,对照组正常培养不干预,辐照组给予4 Gy的辐照量,2 Gy/min,每次辐照2 min,24 h后用实时聚合酶链反应检测细胞中cGAS-STING信号通路相关基因MB21D1、TMEM173、TBK1、IRF3、IRF5、IRF7、IFNAR1、IFNAR2、MX1、MX2、IFIT1、IFIT mRNA。将100μL对数生长期LLC1细胞悬液注射至小鼠腹股沟处皮下,制备双侧皮下荷瘤模型,将成瘤小鼠随机分为对照组、8 Gy组、EGFR-TKI组、8 Gy+EGFR-TKI组,每组3只。对照组不干预;8 Gy组给予8 Gy辐照3次,定位辐照小鼠左边瘤体;EGFR-TKI组给予EGFR-TKI灌胃1 mg/g,给药3次;8 Gy+EGFR-TKI组既进行辐照又给药干预。干预17 d后取小鼠右侧瘤体。用Western blotting法检测肿瘤组织中cGAS、STING、p-STING、Ⅰ型干扰素(IFN-1)蛋白,用免疫组化法检测肿瘤细胞增殖核抗原(ki67)及肿瘤微环境相关蛋白CD4、CD8、表皮生长因子受体(EGFR)、叉头状转录因子p3(Foxp3)、钙网蛋白(CRT)及主要组织相容性复合体(MHC)。结果与对照组比较,辐照组LLC1细胞MB21D1、TMEM173、TBK1、IRF3、IRF5、IFNAR1、IFNAR2、MX1、MX2、IFIT1、IFIT mRNA相对表达量高(P均<0.05),IRF7 mRNA相对表达量低(P<0.05)。干预17 d后,与对照组比较,各干预组肿瘤体积小、肿瘤重量低、ki67蛋白相对表达量低(P均<0.05),且8 Gy+EGFR-TKI组肿瘤体积、肿瘤重量、ki67蛋白相对表达量最低(P均<0.05)。与对照组比较,EGFR-TKI组cGAS、STING、p-STING、IFN-1蛋白相对表达量差异无统计学意义(P均>0.05);8 Gy组STING、IFN-1蛋白相对表达量差异无统计学意义(P均>0.05),cGAS、p-STING蛋白相对表达量高(P均<0.05);8 Gy+EGFR-TKI组cGAS、STING、p-STING、IFN-1蛋白相对表达量高(P均<0.05)。与8 Gy+EGFR-Objective To investigate the effect of epidermal growth factor receptor-tyrosine kinase inhibitor(EGFRTKI)on the abscopal effect of radiotherapy in mice with lung cancer via the cyclic GMP-AMP synthase(cGAS)-stimulator of interferon genes(STING)signaling pathway and the underlying mechanism.Methods Lung cancer LLC1 cells in the logarithmic growth phase were randomly divided into the control group and irradiation group.Cells in the control group were cultured normally without intervention,while cells in the irradiation group were irradiated with 4 Gy(2 Gy/min for 2 min).Twenty-four hours later,the mRNA levels of MB21D1,TMEM173,TBK1,IRF3,IRF5,IRF7,IFNAR1,IFNAR2,MX1,MX2,IFIT1,and IFIT related genes in the cells were determined by real-time PCR(rtPCR).In addition,100μL of LLC1 cell suspension in the logarithmic growth phase was injected subcutaneously into the groin of mice to prepare bilateral subcu⁃taneous tumor-bearing models.The tumor-bearing mice were randomly divided into the control group,8 Gy group,EGFRTKI group,and 8 Gy+EGFR-TKI group,with 3 mice in each group.The tumor-bearing mice in the control group were not intervened.The tumor-bearing mice in the 8 Gy group were radiated at 8 Gy for 3 times,the tumor-bearing mice in the EG⁃FR-TKI group were intragastrically administered 1 mg/g EGFR-TKI for 3 times,and the tumor-bearing mice in the 8 Gy+EGFR-TKI group received both radiation and EGFR-TKI as above.The tumor on the right side of the mice was taken out af⁃ter 17 days of intervention.The levels of cGAS,STING,p-STING,and type I interferon(IFN-1)proteins in the tumor tis⁃sues were determined by Western blotting.Tumor cell proliferation antigen(ki67)and tumor microenvironment-related pro⁃teins CD4,CD8,epidermal growth factor receptor(EGFR),forkhead transcription factor p3(Foxp3),calreticulin(CRT),and major histocompatibility complex(MHC)were detected by immunohistochemistry.Results Compared with the con⁃trol group,LLC1 cells in the radiation group showed significantly higher mRNA expression levels

关 键 词：非小细胞肺癌 表皮生长因子受体酪氨酸激活抑制剂 环鸟苷酸-腺苷酸合成酶-干扰素基因刺激因子信号通路 远隔效应 放射治疗 免疫微环境 小鼠 

分 类 号：R734.2[医药卫生—肿瘤]