肺腺癌患者胸腔积液外泌体蛋白质组学及外泌体蛋白POSTN功能研究  

Study on proteomics of exosomes in pleural effusion from lung adenocarcinoma patients and function of exosomal protein POSTN

作　　者：尹晓翔[1] 刘梦雯 郭颖 赵森[1] YIN Xiaoxiang;LIU Mengwen;GUO Ying;ZHAO Sen(Department of Medical Imaging,the First Affiliated Hospital of Henan University,Henan Kaifeng 475000,China)

机构地区：[1]河南大学第一附属医院医学影像科,河南开封475000

出　　处：《现代肿瘤医学》2025年第5期733-742,共10页Journal of Modern Oncology

基　　金：2021年度河南省医学科技攻关计划联合共建项目(编号:LHGJ20210568)。

摘　　要：目的:挖掘肺腺癌(lung adenocarcinoma,LUAD)患者胸腔积液的潜在外泌体蛋白标志物,探讨外泌体骨膜蛋白(periostin,POSTN)对LUAD细胞增殖和侵袭的影响及其机制,从而为LUAD的早期诊断和治疗提供新思路。方法:首先利用差速超速离心法分离LUAD患者和良性患者的胸腔积液外泌体,随后用电子显微镜、纳米粒径追踪分析、Western Blotting对分离出的外泌体进行鉴定。对提取LUAD患者胸腔积液外泌体与对照组外泌体进行蛋白质谱分析,并通过生物信息学筛选核心差异蛋白,在LUAD A549细胞中过表达和干扰POSTN表达,收集细胞上清外泌体,将其与A549细胞共培养,并进一步通过CCK-8、细胞划痕实验、Transwell实验等体外实验探究外泌体POSTN对肺癌细胞生物学功能的影响。结果:LUAD患者胸腔积液来源的外泌体与良性胸腔积液患者外泌体进行蛋白质组学分析,共鉴定出242种差异表达蛋白,其中表达上调的蛋白有82种,表达下调的蛋白有160种,通过综合生物信息学分析得出了3种关键差异蛋白,进一步通过ROC曲线分析筛选出最有潜力的标志物蛋白POSTN。CCK-8、细胞划痕实验、Transwell实验等体外实验结果显示,用过表达POSTN的外泌体处理A549细胞后,能够对细胞增殖、迁移、侵袭具有促进作用。结论:LUAD患者与良性患者的胸腔积液外泌体中存在差异表达蛋白质,其中外泌体蛋白POSTN可作为LUAD诊断的潜在生物标志物,其在促进LUAD发生和发展中可能发挥重要作用,因此,外泌体POSTN有望成为新生物标志物以及LUAD治疗的潜在靶点。Objective:To explore potential exosomal protein biomarkers in pleural effusion from lung adenocarcinoma(LUAD)patients and investigate the effects and mechanisms of exosomal protein POSTN(periostin)on the proliferation and invasion of LUAD cells,providing new insights for early diagnosis and treatment of LUAD.Methods:Pleural effusions from LUAD patients and benign controls were subjected to differential ultracentrifugation to isolate exosomes.The isolated exosomes were characterized by TEM,Nanosight,and Western Blotting.Proteomic analysis was performed on exosomes extracted from the pleural effusions of LUAD patients and control groups,and core differential proteins were screened using bioinformatics.POSTN expression was overexpressed and knocked down in LUAD A549 cells,and exosomes from these cells were collected and co-cultured with A549 cells.The effects of exosomal POSTN on the biological functions of lung cancer cells were further explored through in vitro experiments such as CCK-8,cell scratch assay,and Transwell assay.Results:Proteomic analysis identified 242 differentially expressed proteins between exosomes from LUAD patients and those from benign pleural effusion patients,with 82 proteins upregulated and 160 downregulated.Comprehensive bioinformatics analysis identified three key differential proteins,and ROC curve analysis further screened POSTN as the most promising biomarker.In vitro experiments including CCK-8,cell scratch assay,and Transwell assay showed that exosomes with overexpressed POSTN promoted A549 cell proliferation,migration,and invasion.Conclusion:Differentially expressed proteins were found in exosomes from pleural effusions of LUAD patients compared to benign controls.Among these,exosomal protein POSTN can be considered as potential biomarker for LUAD diagnosis and may play an important role in promoting the occurrence and development of LUAD.Therefore,exosomal POSTN holds promise as a new biomarker and a potential therapeutic target for LUAD.

关 键 词：肺腺癌 外泌体 胸腔积液 蛋白质组学 POSTN 

分 类 号：R734.2[医药卫生—肿瘤]