电磁脉冲对A549细胞分泌外泌体的影响  

Effects of Electromagnetic Pulses on Exosomes Secretion by A549 Cells

作　　者：侯庆霞 王映梅 曹猛 刘江正 孔德钦 张倩 于卫华[2,3] 安广洲 Qingxia HOU;Yingmei WANG;Meng CAO;Jiangzheng LIU;Deqin KONG;Qian ZHANG;Weihua YU;Guangzhou AN(Department of Pathology,the First Affiliated Hospital of Air Force Medical University,Xi’an 710032,China;Department of Toxicology and Chemical Defense Medicine,Faculty of Preventive Medicine,Air Force Medical University,Xi’an 710032,China;Ministry of Education Key Laboratory of Hazard Assessment and Control in Special Operational Environment,Xi’an 710032,China)

机构地区：[1]空军军医大学第一附属医院病理科,西安710032 [2]空军军医大学军事预防医学系毒理学与防化医学教研室 [3]特殊作业环境危害评估与防治教育部重点实验室

出　　处：《中国肺癌杂志》2024年第12期885-893,共9页Chinese Journal of Lung Cancer

基　　金：国家自然科学基金项目(No.31900888)资助。

摘　　要：背景与目的 大量研究提示电磁脉冲(electromagnetic pulses, EMP)在肿瘤治疗方面具有靶向性强、副作用少和治疗费用低等优点,但其用于肿瘤治疗的最佳参数以及EMP与肿瘤外泌体之间的关系至今仍不清楚。本研究旨在明确不同参数EMP对人非小细胞肺癌A549细胞分泌外泌体数量和内含物微小RNA(microRNA, miRNA)的影响,为EMP临床应用和相关研究提供参考。方法 将A549细胞随机分为对照组和不同强度EMP辐照组,辐照组细胞分别接受场强为400、600和800 kV/m EMP辐照,EMP脉冲数为2000个,重复频率为20 Hz,脉宽为120 ns,每天辐照1次,连续辐照3 d。辐照结束后,收集并鉴定外泌体,台盼蓝染色检测活细胞数量变化,纳米颗粒追踪分析(nanoparticle tracking analysis, NTA)检测外泌体浓度变化,miRNA测序检测外泌体内miRNA丰度变化。结果 与对照组相比,EMP辐照组A549细胞的形态和数量未见明显改变,但400或800 kV/m EMP辐照组外泌体数量明显减少(P<0.05),600 kV/m EMP辐照组外泌体数量明显增多(P<0.05)。各个EMP辐照组与对照组相比,外泌体miRNAs的表达丰度明显不同(P<0.05),其差异丰度miRNAs靶基因富集在不同的细胞信号通路。结论 实验条件下EMP辐照可明显改变A549细胞分泌外泌体的数量和miRNAs的丰度,可进一步影响肿瘤外泌体功能。Background and objective Numerous researches indicated that electromagnetic pulses(EMP)pos-sessed advantages such as strong targeting,minimal side-effects and low treatment cost in tumor therapy,but its optimum parameters for treatment and the relationship between EMP and tumor-derived exosomes remains unclear.This study aims to clarify the effects of EMP with different parameters on the quantity and miRNA(microRNA)of exosomes secreted by human non-small cell lung cancer A549 cells,providing beneficial reference for the clinical application of EMP and related research.Methods A549 cells were randomly divided into control group and different EMP radiation groups with respective intensity of 400,600 and 800 kV/m.EMP was performed with 2000 pulses once,20 Hz of repetition frequency and 120 ns of pulse width.A549 cells were radiated once per day for continuous 3 days.After radiation,exosomes were collected and identified;cell number was measured by trypan blue staining;the concentration of exosomes was measured by nanoparticle tracking analysis(NTA);the abundance of miRNAs was determined by miRNA sequencing.Results Compared with control group,the mor-phology and cell viability of A549 cells in radiation group was not different,but the quantity of exosomes in 400 or 800 kV/m radiation group was significantly decreased(P<0.05),in contrast with obvious increase in 600 kV/m radiation group(P<0.05).The abundance of exosomal miRNAs between control group and each EMP group was obviously different(P<0.05)and target genes of differentially abundant miRNAs enriched in different pathways.Conclusion Under the experimental condition,the quantity and miRNA abundance of exosomes could be changed by EMP radiation,which could further influence the function of tumor-derived exosomes.

关 键 词：肺肿瘤 A549细胞 外泌体 电磁脉冲 MIRNA 

分 类 号：R73[医药卫生—肿瘤]