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

基础医学与临床 ›› 2021, Vol. 41 ›› Issue (11): 1564-1569.

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

朱萌¹, 张宁², 陶伟^3*

1.宁夏医科大学基础医学院,宁夏银川 750004;
宁夏医科大学总医院 2.病理科; 3.消化内科,宁夏银川 750004

收稿日期:2020-11-10 修回日期:2021-04-30 发布日期:2021-10-27
通讯作者: *nyfytaowei@163.com
基金资助:
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)

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

ZHU Meng¹, ZHANG Ning², TAO Wei^3*

1. Basic Medical College, Ningxia Medical University, Yinchuan 750004;
2. Department of Pathology; 3. Department of Gastroenterology, General Hospital,Ningxia Medical University, Yinchuan 750004, China

Received:2020-11-10 Revised:2021-04-30 Published:2021-10-27
Contact: *nyfytaowei@163.com

摘要/Abstract

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。

关键词: 胃癌, 间皮细胞, 外泌体, 微小RNA, Smad7

Abstract: 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 and α-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 with HMrSV5 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 and α-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.

Key words: gastric cancer, mesothelial cells, exosomes, microRNA, Smad7

中图分类号:

R735.2

朱萌, 张宁, 陶伟. 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[J]. 基础医学与临床, 2021, 41(11): 1564-1569.

ZHU Meng, ZHANG Ning, TAO Wei. Effect of exosomes miR-106a derived from human gastric cancer cell line AGS on the phenotype of peritoneal mesothelial cell line HMrSV5[J]. Basic & Clinical Medicine, 2021, 41(11): 1564-1569.

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