人胃癌细胞系AGS外泌体miR-106a对腹膜间皮细胞系HMrSV5表型的影响  

Effect of exosomes miR-106a derived from human gastric cancercell line AGS on the phenotype of peritoneal mesothelial cell line HMrSV5

作　　者：朱萌 张宁[2] 陶伟[3] ZHU Meng;ZHANG Ning;TAO Wei(Basic Medical College, Ningxia Medical University, Yinchuan 750004;Department of Pathology,General Hospital,Ningxia Medical University,Yinchuan 750004,China;Department of Gastroenterology, General Hospital,Ningxia Medical University, Yinchuan 750004, China)

机构地区：[1]宁夏医科大学基础医学院,宁夏银川750004 [2]宁夏医科大学总医院病理科,宁夏银川750004 [3]宁夏医科大学总医院消化内科,宁夏银川750004

出　　处：《基础医学与临床》2021年第11期1564-1569,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(81802416);宁夏自然科学基金(2020AAC02022);宁夏医科大学校级项目(XT2019007)。

摘　　要：目的探讨人胃癌细胞系AGS外泌体及其包裹miR-106a对人腹膜间皮细胞系HMrSV5表型的影响。方法采用离心法分离AGS细胞来源外泌体,与HMrSV5细胞共培养,并加入外泌体分泌抑制剂GW4869。采用qPCR检测miR-106a表达量;EdU检测HMrSV5细胞增殖能力;流式细胞仪检测细胞凋亡。AGS细胞转染miR-106a mimic或inhibitor,共培养后利用Transwell小室法检测HMrSV5细胞迁移;Western blot检测Smad7、TIMP2、MMP2、MMP9、α-SMA蛋白表达。TGF-β处理HMrSV5细胞,共培养后Western blot检测Smad7、Smad2/3、p-Smad2/3蛋白表达。结果AGS细胞高表达外泌体miR-106a(P<0.01);与HMrSV5细胞共培养后明显抑制其增殖,促使早期凋亡率增加(P<0.05)。转染miR-106a mimic后,AGS细胞源外泌体促进HMrSV5细胞迁移(P<0.001),同时Smad7、TIMP2表达下降,MMP2、MMP9、α-SMA表达升高,阻断miR-106a传递后这一作用减弱。TGF-β处理后,Smad7表达减弱,Smad2/3、p-Smad2/3表达升高。结论AGS细胞来源外泌体能够促进HMrSV5细胞凋亡、抑制增殖、增强迁移,可能与转运miR-106a靶向Smad7/TIMP2有关。Objective To investigate the effect of exosomes derived from human gastric cancer cell line AGS and miR-106a on the phenotype of peritoneal mesothelial cells line HMrSV5.Methods Exosomes from AGS cells were isolated and incubated with HMrSV5 cells.Then exosome secretion inhibitor GW4869 was added.miR-106a expression was detected by qPCR,HMrSV5 cells proliferation and apoptosis were detected by EdU and flow cytometry technology.After the AGS cells were transfected with miR-106a mimic or inhibitor,the migration of HMrSV5 cells was detected by Transwell assay,and the expression of Smad7,TIMP2,MMP2,MMP9 andα-SMA was detected by Western blot.After treatment with TGF-β,the expression of Smad7,Smad2/3 and p-Smad2/3 was detected by Western blot.Results The exosomes of AGS cells highly expressed miR-106a(P<0.01),and incubation withHMrSV5 cells significantly inhibited its proliferation and promoted the early apoptosis rate(P<0.05).After transfection of miR-106a mimic,the exosomes from AGS cells promoted the migration of HMrSV5 cells(P<0.001).Meanwhile,the expression of Smad7 and TIMP2 was decreased,but the expression of MMP2,MMP9 andα-SMA was increased.After treatment with TGF-β,the expression of Smad7 decreased but the expression of Smad2/3 and p-Smad2/3 increased.Conclusions Exosomes derived from human gastric cancer AGS cells can promote apoptosis,inhibit proliferation and enhance migration of human peritoneal mesothelial HMrSV5 cells,which may be related to the transport of miR-106a targeting Smad7 and TIMP2.

关 键 词：胃癌 间皮细胞 外泌体 微小RNA SMAD7 

分 类 号：R735.2[医药卫生—肿瘤]