胃癌肿瘤相关巨噬细胞内SHP2通过抑制STAT3-TGFβ通路进而减轻PD-L1表达  

Tumor-associated macrophage tyrosine phosphatase SHP2 reduces PD-L1 expression in gastric cancer by inhibiting STAT3-TGFβpathway

作　　者：李帅 石玉[1] 王常昊 艾冬梅[1] 张卜瑗[1] Li Shuai;Shi Yu;Wang Changhao;Ai Dongmei;Zhang Buyuan(Department of Medical Oncology,The Second People’s Hospital of Hengshui,Hengshui 053000,China)

机构地区：[1]衡水市第二人民医院肿瘤内科,衡水053000

出　　处：《解剖学杂志》2024年第5期398-403,463,共7页Chinese Journal of Anatomy

摘　　要：目的:探究肿瘤相关巨噬细胞内含Src同源2结构域蛋白酪氨酸磷酸酶(SHP2)对胃癌内细胞程序性死亡-配体1(PD-L1)表达的影响及其机制。方法:将人单核/巨噬细胞THP-1细胞、人胃癌细胞系SGC-7901细胞培养后,THP-1细胞诱导分化为巨噬细胞,采用慢病毒感染的方法构建稳定敲低和过表达SHP2基因的THP-1细胞,与SGC-7901细胞非接触式共培养,将其分为对照(NC)组和SHP2-shRNA组、NC组和SHP2-mimic组、Stattic(STAT3抑制剂)+NC组和Stattic+SHP2-shRNA组。利用超速离心法提取THP-1细胞上清液外泌体,免疫印迹检测THP-1细胞中STAT3-TGFβ通路相关蛋白及外泌体CD9和TGFβ蛋白表达情况,免疫印迹检测SGC-7901细胞STAT3-TGFβ通路相关蛋白及PD-L1、p-STAT3、IL-10、PTEN和p-PI3K蛋白表达情况,CCK-8检测SGC-7901细胞增殖活性,Transwell实验检测细胞迁移和侵袭能力。结果:THP-1细胞中,SHP2-shRNA组SHP2蛋白表达水平显著低于NC组,p-STAT3、IL-10和TGFβ及外泌体中CD9和TGFβ蛋白表达水平显著高于NC组;SHP2-mimic组SHP2蛋白表达水平显著高于NC组,p-STAT3、IL-10和TGFβ及外泌体中CD9和TGFβ蛋白表达水平显著低于NC组。共培养的SGC-7901细胞中,SHP2-shRNA组SHP2和PTEN蛋白表达水平显著低于NC组,p-STAT3、TGFβ、PD-L1和p-PI3K蛋白表达水平及细胞增殖活性、迁移和侵袭能力显著高于NC组;Stattic干预后,SHP2蛋白表达水平未被影响,PTEN蛋白表达水平增加,p-STAT3、TGFβ、PD-L1和p-PI3K蛋白表达水平及细胞增殖活性、迁移和侵袭能力降低,Stattic+NC组和Stattic+SHP2-shRNA组之间差异无统计学意义。结论:肿瘤相关巨噬细胞SHP2通过抑制胃癌细胞内STAT3-TGFβ通路活性抑制PD-L1和p-PI3K表达,进而抑制胃癌细胞增殖活性及迁移和侵袭能力,从而延缓胃癌进展。Objective:To explore the effect of tumor-associated macrophage tyrosine phosphatase SHP2 on PDL1 expression in gastric cancer and preliminarily elucidate its mechanism.Methods:After human monocyte/macrophage THP-1 cells and human gastric cancer cell line SGC-7901 cells were cultured,THP-1 cells were induced to differentiate into macrophages,and THP-1 cells with stable knockdown and overexpression of SHP2 gene were constructed by lentivirus infection.The infected THP-1 cells were co-cultured with SGC-7901 cells in a non-contact mode,and they were divided into the NC group and SHP2-shRNA group,NC group and SHP2-mimic group,and Stattic(STAT3 inhibitor)+NC group and Stattic+SHP2-shRNA group.Supernatant exosomes of THP-1 cells were extracted by ultracentrifugation,and the expressions of STAT3-TGFβpathwayrelated proteins,exosomes CD9,and TGFβproteins in THP-1 cells were detected by Western blotting.Western blotting was also used to detect the expression of STAT3-TGFβpathway-related proteins and PD-L1,PTEN,and p-PI3K proteins in SGC-7901 cells.CCK-8 assay was used to detect the proliferation activity of SGC-7901 cells.Transwell assay was used to detect the migration and invasion abilities of SGC-7901 cells.Results:The expression level of SHP2 protein in the THP-1 cells of the SHP2-shRNA group was significantly lower than that in the NC group,and the expression levels of p-STAT3,IL-10,TGFβ,as well as CD9 and TGFβproteins in exosomes in the SHP2-shRNA group were significantly higher than those in the NC group.The expression level of SHP2 protein in the SHP2-mimic group was significantly higher than that in the NC group,and the expression levels of p-STAT3,IL-10,TGFβ,as well as CD9 and TGFβproteins in exosomes were significantly lower than those in the NC group.In SGC-7901 cells during co-culture,the protein expression levels of SHP2 and PTEN in the SHP2-shRNA group were significantly lower than those in the NC group,while the protein expression levels of p-STAT3,TGFβ,PD-L1,and p-PI3K,as well as the cell proliferat

关 键 词：肿瘤相关巨噬细胞 含Src同源2结构域蛋白酪氨酸磷酸酶 信号转导和转录激活因子3-转化生长因子β信号通路 胃癌 细胞程序性死亡-配体1 肿瘤微环境 

分 类 号：R392.32[医药卫生—免疫学]