过表达M2型肿瘤相关巨噬细胞TIA1基因通过调控PI3K/AKT信号通路抑制胃癌细胞侵袭和迁移  

Overexpression of TIA1 gene in M2-type tumor-associated macrophages inhibited invasion and migration of gastric cancer cells by regulating the PI3K/AKT signaling pathway

作　　者：钟裕昌 阮笃激 戴霖 王彪 曲颜 杨雪峰 ZHONG Yuchang;RUAN Duji;DAI Lin;WANG Biao;QU Yan;YANG Xuefeng(Department of Gastrointes-tinal Surgery,the Second Affiliated Hospital of Zunyi Medical University,Zunyi 563006,China;Department of Gastrointestinal Surgery,Affiliated Hospital of Zunyi Medical University,Zunyi 563006,China)

机构地区：[1]遵义医科大学第二附属医院胃肠外科,遵义563006 [2]遵义医科大学附属医院胃肠外科,遵义563006

出　　处：《中国免疫学杂志》2024年第8期1658-1664,共7页Chinese Journal of Immunology

基　　金：贵州省科技计划项目(黔科合基础[2020]1Y335)。

摘　　要：目的:探讨过表达M2型肿瘤相关巨噬细胞(M2-TAMs)T淋巴细胞内抗原1(TIA1)基因对胃癌BGC-823细胞侵袭和迁移的影响及其机制。方法:从胃癌组织中提取原代TAMs,采用IL-4和IL-13刺激TAMs诱导分化成M2-TAMs,再将TIA1基因过表达质粒(oe-TIA1)及其空载质粒(Vector)转染至M2-TAMs中,采用qRT-PCR和Western blot检测细胞中TIA1 mRNA和蛋白表达水平;流式细胞术检测细胞中CD206和CD163表达水平;ELISA检测细胞培养上清液中IL-10、TGF-β、VEGF-A和Arg-1水平。通过Transwell共培养体系将转染后的M2-TAMs与胃癌BGC-823细胞共培养,并联用PI3K激动剂740Y-P进行干预,采用划痕实验检测细胞迁移能力;Transwell实验检测细胞侵袭能力;Western blot检测细胞中PI3K、p-PI3K、AKT、p-AKT、MMP-2和MMP-9等蛋白表达水平。结果:①与原代TAMs比较,M2-TAMs中CD206和CD163表达水平及细胞培养上清液中IL-10、TGF-β、VEGF-A和Arg-1水平均显著升高(P<0.05),而TIA1 mRNA和蛋白表达水平显著降低(P<0.05)。TIA1基因过表达可显著降低M2-TAMs中CD206和CD163表达水平及细胞培养上清液中IL-10、TGF-β、VEGF-A和Arg-1水平(P<0.05)。②M2-TAMs中过表达TIA1基因可显著降低BGC-823细胞迁移和侵袭能力及p-PI3K/PI3K、p-AKT/AKT、MMP-2和MMP-9等蛋白表达水平(P<0.05)。③740Y-P可显著逆转M2-TAMs中过表达TIA1基因对BGC-823细胞迁移、侵袭及PI3K/AKT信号通路的抑制作用。结论:过表达M2-TAMs中TIA1基因表达可通过阻断PI3K/AKT信号通路影响胃癌细胞侵袭和迁移。Objective:To investigate the effect of overexpression of T cell restricted intracellular antigen 1(TIA1)gene in M2-type tumor-associated macrophages(M2-TAMs)on invasion and migration of gastric cancer cells and its mechanism.Methods:Primary TAMs were extracted from gastric cancer tissues and induced to differentiate into M2-TAMs using IL-4 and IL-13.The TIA1 overexpression plasmid(oe-TIA1)and its empty vector were transfected into M2-TAMs.The expression levels of TIA1 mRNA and protein were detected by qRT-PCR and Western blot.The expression levels of CD206 and CD163 were detected by flow cytometry.The levels of IL-10,TGF-β,VEGF-A and Arg-1 in cell culture supernatant were detected by ELISA.The transfected M2-TAMs were co-cultured with gastric cancer BGC-823 cells by Transwell co-cultivation system,and PI3K agonist 740Y-P was used to intervene in parallel.The cell migration ability was detected by scratch assay.Transwell assay was used to detect cell invasion ability.Protein expression levels of PI3K,p-PI3K,AKT,p-AKT,MMP-2 and MMP-9 in the cells were detected by Western blot.Results:①Compared with primary TAMs,the levels of CD206 and CD163 expression in M2-TAMs cells and IL-10,TGF-β,VEGF-A and Arg-1 in cell culture supernatant were significantly increased(P<0.05),while the expression levels of TIA1 mRNA and protein were significantly decreased(P<0.05).Overexpression of TIA1 gene significantly decreased the expression levels of CD206 and CD163 in M2-TAMs and IL-10,TGF-β,VEGF-A and Arg-1 in cell culture supernatant(P<0.05).②Overexpression of TIA1 gene in M2-TAMs could significantly reduce the migration and invasion ability and the expression levels of p-PI3K/PI3K,p-AKT/AKT,MMP-2 and MMP-9 proteins in BGC-823 cells(P<0.05).③740Y-P could significantly reverse the inhibitory effects of overexpression of TIA1 gene in M2-TAMs on migration,invasion and PI3K/AKT signaling pathway of BGC-823 cells.Conclusion:Overexpression of TIA1 gene in M2-TAMs can affect the invasion and migration of gastric cancer cells by bloc

关 键 词：M2型肿瘤相关巨噬细胞 T淋巴细胞内抗原1 胃癌细胞 PI3K/AKT信号通路 

分 类 号：R735.2[医药卫生—肿瘤]