肺腺癌IL⁃22表达水平及其促进肺腺癌转移机制的实验研究  

Experimental study on expression level of IL‐22 in lung adenocarcinoma and its mechanism in promoting lung adenocarcinoma metastasis

作　　者：刘蔚然 吴新艺 王长利 张彬 Liu Weiran;Wu Xinyi;Wang Changli;Zhang Bin(Department of Anesthesia,Tianjin Medical University Cancer Institute&Hospital,National Clinical Research Center for Cancer,Tianjin Key Laboratory of Cancer Prevention and Therapy,Tianjin's Clinical Research Center for Cancer,Tianjin 300060,China;Department of Lung Cancer,Tianjin Medical University Cancer Institute&Hospital,National Clinical Research Center for Cancer,Tianjin Key Laboratory of Cancer Prevention and Therapy,Tianjin's Clinical Research Center for Cancer,Tianjin 300060,China)

机构地区：[1]天津医科大学肿瘤医院麻醉科国家恶性肿瘤临床医学研究中心天津市“肿瘤防治”重点实验室天津市恶性肿瘤临床医学研究中心,天津300060 [2]天津医科大学肿瘤医院肺部肿瘤科国家恶性肿瘤临床医学研究中心天津市“肿瘤防治”重点实验室天津市恶性肿瘤临床医学研究中心,天津300060

出　　处：《肿瘤研究与临床》2025年第3期177-185,共9页Cancer Research and Clinic

基　　金：天津市医学重点学科(专科)建设项目(TJYXZDXK⁃010A)。

摘　　要：目的探讨肺腺癌白细胞介素(IL)⁃22的表达及其对肺腺癌转移的影响和可能机制。方法回顾性收集2023年1月至6月天津医科大学肿瘤医院27例肺腺癌患者手术切除的癌组织和配对癌旁(距肿瘤边缘>2 cm)正常组织,流式细胞术检测两组织中T细胞IL⁃22表达水平,比较癌和癌旁组织间、是否淋巴结转移患者肺癌组织间T细胞IL⁃22表达水平;回顾性收集同期另外6例肺腺癌患者的癌组织和配对的癌旁正常组织,蛋白质印迹法检测两组织中IL⁃22受体IL⁃22RA1的表达水平。收集癌症基因组图谱(TCGA)数据库和基因表达综合(GEO)数据库的3个数据集中肺腺癌患者癌组织IL⁃22RA1转录组数据,利用R软件survminer包选取反映生存关系的IL⁃22RA1的最佳临界值,并依此将患者分为IL⁃22RA1高、低两组,用survival包绘制总生存曲线,采用log⁃rank检验进行组间比较。用重组IL⁃22处理人肺腺癌A549细胞、小鼠肺腺癌LLC细胞,均以未经IL⁃22处理的细胞为对照;采用Transwell实验检测各组迁移细胞数;采用蛋白质印迹法检测各组细胞ERK、AKT、STAT信号通路相关蛋白及基质金属蛋白酶9(MMP⁃9)、上皮型钙黏蛋白(E⁃cad)表达水平,检测加入STAT3抑制剂C188⁃9、AKT抑制剂MK⁃2206、ERK抑制剂SCH772984后A549细胞和LLC细胞上述蛋白表达水平。利用10只C57BL/6小鼠构建LLC细胞的肺转移模型,简单随机法将小鼠平均分为实验组和对照组,分别隔日注射1次IL⁃22中和抗体(50µg/只)和无中和作用的对照抗体(50µg/只),第10天处死并解剖小鼠,计数肺转移结节,转移肺组织经HE染色并计数其中转移灶,采用流式细胞术检测由转移肺组织制备的单细胞中CD4^(+)T细胞、CD8^(+)T细胞占免疫细胞的比例。结果流式细胞术检测显示,临床收集的27例肺腺癌组织IL⁃22^(+)CD4^(+)T细胞在T细胞(由CD3、CD45标记)中的占比[M(Q1,Q3)]高于癌旁正常组织[0.28%(0.04%,1.00%)比0.01%Objective To investigate the expression of interleukin(IL)-22 in lung adenocarcinoma and its effect and possible mechanism for lung adenocarcinoma metastasis.Methods The cancer tissues and paired adjacent normal tissues(>2 cm from the tumor edge)surgically removed from 27 lung adenocarcinoma patients in Tianjin Medical University Cancer Institute&Hospital from January to June 2023 were retrospectively collected.Flow cytometry was used to detect the expression levels of IL-22 in T cells of all tissues,and the expression levels of IL-22 in T cells were compared between cancer and adjacent tissues,as well as between lung cancer tissues of patients with and without lymph node metastasis.The cancer tissues and paired adjacent normal tissues were retrospectively collected from 6 patients with lung adenocarcinoma during the same period,and the expression level of IL-22 receptor IL-22RA1 in the tissues was detected by Western blotting.IL-22RA1 transcriptome data from cancer tissues of lung adenocarcinoma patients in 3 datasets in The Cancer Genome Atlas(TCGA)database and the Gene Expression Omnibus(GEO)database were collected.Using the R software survminer package to select the optimal critical value of IL-22RA1 that reflected the survival relationship,and patients were divided into high and low IL-22RA1 groups based on this.The survival package was used to draw the overall survival curve and log-rank test was performed for inter group comparison.Recombinant IL-22 was used to treat human lung adenocarcinoma A549 cells and mouse lung adenocarcinoma LLC cells,with cells not treated with IL-22 as controls;Transwell assay was used to detect the number of migrating cells in each group;Western blotting was used to detect the expression levels of ERK,AKT and STAT signaling pathwaysrelated proteins,matrix metalloproteinase 9(MMP-9)and epithelial cadherin(E-cad)in each group of cells.The expression levels of these proteins in A549 cells and LLC cells were also measured after the addition of STAT3 inhibitor C188-9,AKT inhibitor MK

关 键 词：肺肿瘤 腺癌 白细胞介素22 肿瘤转移 肿瘤微环境 

分 类 号：R734.2[医药卫生—肿瘤]