胃癌来源间充质干细胞通过JAK2/STAT3信号通路调控胃癌微环境中巨噬细胞向M2亚型极化  

Gastric cancer-derived mesenchymal stem cells regulate the M2 polarization of macrophages within gastric cancer microenvironment via JAK2/STAT3 signaling pathway

作　　者：李伟[1] 赵绍林[2] 郑萍[2] 史沛芹[2] 周颖[2] 张婷[1] 霍娟[1] 杨晋[2] Li Wei;Zhao Shaolin;Zheng Ping;Shi Peiqin;Zhou Ying;Zhang Ting;Huo Juan;Yang Jin(Center Laboratory,the First People′s Hospital of Lianyungang,Lianyungang 222001,China;Clinical Laboratory,the First People′s Hospital of Lianyungang,Lianyungang 222001,China)

机构地区：[1]连云港市第一人民医院中心实验室,连云港222001 [2]连云港市第一人民医院检验科,连云港222001

出　　处：《中华肿瘤杂志》2022年第7期728-736,共9页Chinese Journal of Oncology

基　　金：国家自然科学基金青年科学基金项目(81402280);江苏省自然科学基金面上项目(BK20161296);江苏省博士后科研资助计划项目(1501079A)。

摘　　要：目的探讨胃癌微环境中肿瘤来源间充质干细胞对巨噬细胞M2亚型极化的调节作用及机制。方法胃癌及癌旁正常组织来源于2018年于连云港市第一人民医院行胃癌切除的患者(4例)。将THP-1诱导分化来源的巨噬细胞与胃癌来源间充质干细胞(GC-MSCs)进行体外共培养,细胞分为对照组(Mac组)、细胞上清处理组(Mac+GC-MSC-CM组)、Mac+GC-MSC-TW组、Mac+IL-6-blocked-GC-MSC-CM组、Mac+IL-8-blocked-GC-MSC-CM组和Mac+IL-6/IL-8-blocked-GC-MSC-CM组。采用实时荧光定量聚合酶链反应、流式细胞术和Luminex液态芯片技术,分别评价巨噬细胞中M2亚型相关基因、细胞表面标记蛋白以及细胞因子谱的表达情况。利用Luminex液态芯片技术检测并筛选GC-MSCs中可能发挥诱导巨噬细胞向M2亚型极化作用的关键细胞因子,并采用中和抗体试验进行验证。Western blot法检测GC-MSCs不同处理条件下,巨噬细胞中M2亚型极化相关信号通路蛋白表达情况。结果Mac+GC-MSC-CM组巨噬细胞中M2亚型相关基因Ym-1和Fizz-1表达水平分别为1.53±0.32和13.22±1.05,均高于Mac组(分别为1.00±0.05和1.21±0.38,均P<0.05),iNOS基因表达水平(0.60±0.41)低于Mac组(1.06±0.38,P=0.023)。Mac+GC-MSC-TW组巨噬细胞中Ym-1和Fizz-1基因表达水平分别为1.47±0.09和13.16±2.77,均高于Mac组(分别为1.00±0.05和1.21±0.38,均P<0.05),iNOS基因表达水平(0.56±0.03)低于Mac组(1.06±0.38,P=0.026)。Mac+GC-MSC-CM组和Mac+GC-MSC-TW组巨噬细胞中CD163+和CD204^(+)细胞比例分别为3.80%和4.40%,均高于Mac组(0.60%)。Mac+GC-MSC-CM组巨噬细胞中白细胞介素10(IL-10)、IL-6、单核细胞趋化蛋白1和血管内皮生长因子的表达水平分别为(592.60±87.52)pg/ml、(1346.80±64.70)pg/ml、(11256.00±29.03)pg/ml和(1463.90±66.67)pg/ml,均高于Mac组[分别为(41.03±2.59)pg/ml、(17.35±1.79)pg/ml、(5213.30±523.71)pg/ml和(267.12±12.06)pg/ml,均P<0.05],肿瘤坏死因子α、IP-10、RANTES和巨噬细胞炎症蛋白1α�Objective To investigate the role and mechanism of tumor-derived mesenchymal stem cells in regulating the M2 polarization of macrophages within gastric cancer microenvironment.Methods Gastric cancer tissues and the adjacent non-cancerous tissues were collected from patients underwent gastric cancer resection in the First People′s Hospital of Lianyungang during 2018.In our study,THP-1-differentiated macrophages were co-cultured with gastric cancer-derived mesenchymal stem cells(GC-MSCs).Then,the M2 subtype-related gene,the markers expressed on cell surface and the cytokine profile were analyzed by real-time quantitative reverse transcription polymerase chain reaction(qRT-PCR),flow cytometry and Luminex liquid chip,respectively.The key cytokines mediating the inducing effect of GC-MSCs on macrophage polarization into the M2 subtype were detected and screened by Luminex liquid chip,which were further confirmed by the neutralizing antibody test.The expressions of macrophage proteins involved in M2 polarization-related signaling pathways under the different co-culture conditions of GC-MSCs were detected by western blot.Results In Mac+GC-MSC-culture medium(CM)group,the expression levels of Ym-1 and Fizz-1(1.53±0.32 and 13.22±1.05,respectively),which are markers for M2 subtype,were both significantly higher than those of Mac group(1.00±0.05 and 1.21±0.38,respectively,P<0.05).The level of iNOS in Mac+GC-MSC-CM group(0.60±0.41)was significantly lower than that of Mac group(1.06±0.38,P=0.023).In Mac+GC-MSC-Transwell(TW)group,the expression levels of Ym-1 and Fizz-1(1.47±0.09 and 13.16±2.77,respectively)were both significantly higher than those of Mac group(1.00±0.05 and 1.21±0.38,respectively,P<0.05).The level of iNOS in Mac+GC-MSC-CM group(0.56±0.03)was significantly lower than that of Mac group(1.06±0.38,P=0.026).The ratios of CD163+/CD204^(+)cells in Mac+GC-MSC-CM and Mac+GC-MSC-TW groups(3.80%and 4.40%,respectively)were both remarkably higher than that of Mac group(0.60%,P<0.05).The expression levels of I

关 键 词：胃肿瘤 间充质干细胞 巨噬细胞 极化 

分 类 号：R735.2[医药卫生—肿瘤]