缺氧外泌体传递miR-199a-5p对胃癌细胞SGC-7901迁移和侵袭的影响  

Effects of miR-199a-5p Delivered by Hypoxic Exosomes on the Migration and Invasion of Gastric Cancer Cells SGC-7901

作　　者：周辉 沈伟锋 邵平扬 ZHOU Hui;SHEN Wei-feng;SHAO Ping-yang(Department of Clinical Laboratory,the FirstHospital of Jiaxing,Jiaxing 314001,Zhejiang,China)

机构地区：[1]嘉兴市第一医院检验科,浙江嘉兴314001

出　　处：《医学信息》2021年第1期78-82,86,共6页Journal of Medical Information

基　　金：嘉兴市科技计划(编号:2018AD32059);2019年嘉兴市医学重点学科-临床检验诊断学(创新学科)(编号:2019-cx-03)。

摘　　要：目的探讨缺氧环境下胃癌细胞SGC-7901源性外泌体携带miRNA对常氧胃癌细胞迁移和侵袭的作用。方法使用含CoCl2培养基培养SGC-7901细胞模拟细胞缺氧,将缺氧源性的外泌体与常氧条件下SGC-7901细胞共培养,采用划痕实验和侵袭实验检测缺氧胃癌细胞源性外泌体对常氧胃癌细胞迁移和侵袭能力的影响,采用高通量测序筛查缺氧与常氧外泌体中差异表达的miRNA;并调节缺氧胃癌细胞上述差异表达miRNA水平,提取外泌体与常氧胃癌细胞共培养,采用划痕实验和侵袭实验检测外泌体携带miRNA对常氧胃癌细胞迁移和侵袭能力的影响。结果透射电子显微镜下显示,常氧和缺氧SGC-7901细胞分泌的外泌体均呈直径为50~150nm圆形或椭圆形的杯状结构。Western免疫印记分析,常氧和缺氧SGC-7901细胞分泌外泌体均表达CD63和Tsg101蛋白标志物。划痕及侵袭实验显示,缺氧处理SGC-7901细胞分泌的外泌体共培养的常氧SGC-7901细胞划痕闭合率、移动到transwell小室下方的数量高于常氧SGC-7901细胞分泌的外泌体共培养的常氧SGC-7901细胞(P<0.05)。测序和RT-PCR检测显示,缺氧外泌体中miR-199a-5p的表达高于常氧外泌体(P<0.05)。划痕及侵袭实验显示,导入miR-199a-5p mimics细胞来源的外泌体共培养的常氧SGC-7901细胞划痕闭合率、移动到transwell小室下方的数量高于导入miR-199a-5p mimicsNC细胞来源的外泌体共培养的常氧SGC-7901细胞(P<0.05)。结论缺氧微环境可能会刺激胃癌细胞产生富含miR-199a-5p的外泌体,并将其输送至周围常氧胃癌细胞以促进其迁移和侵袭。Objective To explore the effect of gastric cancer cell SGC-7901-derived exosomes carrying miRNA on the migration and invasion of normoxic gastric cancer cells under hypoxia.Methods SGC-7901 cells were cultured with CoCl2 medium to simulate cell hypoxia.Hypoxic exosomes were co-cultured with SGC-7901 cells under normoxia.Scratch test and invasion test were used to detect the source of hypoxic gastric cancer cells The effect of sex exosomes on the migration and invasion of normoxic gastric cancer cells,using high-throughput sequencing to screen the differentially expressed miRNAs in hypoxic and normoxic exosomes;And adjust the level of the above-mentioned differentially expressed miRNA in hypoxic gastric cancer cells,extract exosomes and co-culture with normoxic gastric cancer cells,use scratch test and invasion experiment to detect the effect of exosomes carrying miRNA on the migration and invasion ability of normoxic gastric cancer cells.Results The transmission electron microscope showed that the exosomes secreted by the normoxic and hypoxic SGC-7901 cells were round or elliptical cup-shaped structures with a diameter of 50-150 nm.Western immunoblotting analysis showed that both normoxic and hypoxic SGC-7901 cells secreted exosomes express CD63 and Tsg101 protein markers.Scratch and invasion experiments showed that the normal oxygen SGC-7901 cells co-cultured with exosomes secreted by hypoxia treated SGC-7901 cells had a higher scratch closure rate and the number of moving to the bottom of the transwell chamber than those secreted by normoxic SGC-7901 cells.Normal oxygen SGC-7901 cells co-cultured with exosomes(P<0.05).Sequencing and RT-PCR showed that the expression of miR-199a-5p in hypoxic exosomes was higher than that of normoxic exosomes(P<0.05).Scratch and invasion experiments showed that the normal oxygen SGC-7901 cells co-cultured with exosomes derived from miR-199a-5p mimics cells had a scratch closure rate,the number of moving to the bottom of the transwell chamber was higher than that of normoxic SGC-

关 键 词：胃癌 外泌体 缺氧 迁移 侵袭 miR-199a-5p 

分 类 号：R735.2[医药卫生—肿瘤]