肺腺癌源外泌体过表达miR-197-3p影响肿瘤血管生成的作用和机制  

Role of miR-197-3p overexpression in exosomes derived from lung adenocarcinoma in influencing tumor angiogenesis and the underlying mechanism

作　　者：唐文辉 王冬梅 詹紫娥 宛太 Tang Wenhui;Wang Dongmei;Zhan Zi′e;Wan Tai(Department of Pulmonary and Critical Care Medicine,Shenzhen Qianhai Shekou Free Trade Zone Hospital,Shenzhen 518067,China)

机构地区：[1]深圳市前海蛇口自贸区医院呼吸与危重症医学科,深圳518067

出　　处：《国际呼吸杂志》2025年第3期207-215,共9页International Journal of Respiration

基　　金：2022深圳市科技创新委员会资助项目(JCYJ20220530151612028)。

摘　　要：目的探讨肺腺癌源外泌体过表达miR-197-3p影响肿瘤血管生成的作用和机制。方法本研究为实验研究,采用非随机抽样的方法选取2021年10月至2023年10月在深圳市前海蛇口自贸区医院接受外科手术的20例肺腺癌患者为研究对象。于术前采集所有患者的血液样本,并于术中收集所有患者的肺腺癌组织和癌旁正常肺组织样本,另外采集该院同期20名健康对照者的血液样本。提取肺腺癌患者和健康对照者的血浆外泌体、肺腺癌患者的肺腺癌组织和癌旁正常肺组织外泌体、肺腺癌细胞系(A549、PC9、HTB182、H1299、H2009、LTEP-a、H446、H460、BEAS2b)和正常对照细胞系(HBE)外泌体。根据共培养血浆外泌体的不同将人脐静脉血管内皮细胞(HUVECs)分为肿瘤外泌体组和空白外泌体组(分别加入肺腺癌患者血浆外泌体、健康对照者血浆外泌体),根据共培养肺腺癌细胞系外泌体的不同将HUVECs分为高转移潜能组和低转移潜能组(分别加入H1299外泌体、LTEP-a外泌体),进行体外血管形成实验。采用简单随机抽样法从上述20例肺腺癌患者和20名健康对照者中分别抽取6例肺腺癌患者和6名健康对照者,通过高通量测序技术对肺腺癌患者和健康对照者血浆外泌体中包含的微RNA(miRNA)进行测序,获得肺腺癌患者和健康对照者的miRNA差异表达谱,并确定miR-197-3p为本研究中的主要miRNA。采用实时荧光定量聚合酶链反应检测肺腺癌组织与癌旁正常肺组织(组织和外泌体)、肺腺癌细胞系与正常对照细胞系(细胞和外泌体)中miR-197-3p的表达水平。采用慢病毒构建稳定敲除miR-197-3p的H1299细胞系和稳定过表达miR-197-3p的LTEP-a细胞系,将其中提取的外泌体分别与HUVECs共培养,将HUVECs分为H1299敲除组和LTEP-a过表达组;将正常H1299细胞系、LTEP-a细胞系提取的外泌体分别与HUVECs共培养,将HUVECs分为H1299对照组和LTEP-a对照组,进行体外血管�ObjectiveTo explore the role of miR-197-3p overexpression in exosomes derived from lung adenocarcinoma in influencing tumor angiogenesis and the underlying mechanism.MethodsThis was an experimental study.Using non-random sampling,20 patients with lung adenocarcinoma who underwent surgical operations in Shenzhen Qianhai Shekou Free Trade Zone Hospital from October 2021 to October 2023 were selected as the research subjects.Blood samples of all patients were preoperatively collected,and lung adenocarcinoma tissues and adjacent normal lung tissue samples were intraoperatively collected.In addition,blood samples of 20 healthy controls in the same period were collected.Plasma exosomes of patients with lung adenocarcinoma and healthy controls,exosomes of lung adenocarcinoma tissues and adjacent normal lung tissues of patients with lung adenocarcinoma,and exosomes of lung adenocarcinoma cell lines(A549,PC9,HTB182,H1299,H2009,LTEP-a,H446,H460,BEAS2b)and normal control cell line(HBE)were extracted.Human umbilical vein endothelial cells(HUVECs)were divided into the tumor exosome group(treatment of plasma exosomes derived from lung adenocarcinoma)and the blank exosome group(treatment of plasma exosomes derived from healthy controls)according to the treatment of different co-cultured plasma exosomes.Moreover,HUVECs were divided into the high metastatic potential group(treatment of exosomes derived from H1299 cells)and the low metastatic potential group(treatment of exosomes derived from LTEP-a cells)according to the treatment of different co-cultured exosomes of lung adenocarcinoma cell lines,following in vitro angiogenesis experiments.By simple random sampling,6 patients with lung adenocarcinoma and 6 healthy controls were respectively selected from the above-mentioned 20 lung adenocarcinoma patients and 20 healthy controls.High-throughput sequencing technology was used to sequence the microRNA(miRNA)contained in the plasma exosomes of patients with lung adenocarcinoma and healthy controls,thus obtaining differentially expr

关 键 词：肺腺癌 外泌体 微RNAS 胰岛素样生长因子结合蛋白质3 肿瘤血管生成 

分 类 号：R734.2[医药卫生—肿瘤]