Hou Qingxia, Wang Yingmei, Cao Meng, Liu Jiangzheng, Kong Deqin, Zhang Qian, Yu Weihua, An Guangzhou
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.
Zhongguo Fei Ai Za Zhi. 2024 Dec 20;27(12):885-893. doi: 10.3779/j.issn.1009-3419.2024.106.34.
Numerous researches indicated that electromagnetic pulses (EMP) possessed 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.
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.
Compared with control group, the morphology 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.
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.
众多研究表明，电磁脉冲（EMP）在肿瘤治疗中具有靶向性强、副作用小、治疗成本低等优势，但其最佳治疗参数以及与肿瘤来源外泌体之间的关系尚不清楚。本研究旨在阐明不同参数的电磁脉冲对人非小细胞肺癌A549细胞分泌外泌体的数量及微小RNA（miRNA）的影响，为电磁脉冲的临床应用及相关研究提供有益参考。
将A549细胞随机分为对照组和不同强度（400、600和800 kV/m）的电磁脉冲辐射组。电磁脉冲以2000个脉冲单次发射，重复频率为20 Hz，脉冲宽度为120 ns。每天对A549细胞进行一次辐射，连续辐射3天。辐射后，收集并鉴定外泌体；通过台盼蓝染色法检测细胞数量；采用纳米颗粒跟踪分析（NTA）测定外泌体浓度；通过miRNA测序确定miRNA的丰度。
与对照组相比，辐射组A549细胞的形态和细胞活力无差异，但400或800 kV/m辐射组的外泌体数量显著减少（P<0.05），而600 kV/m辐射组则明显增加（P<0.05）。对照组与各电磁脉冲组外泌体miRNA的丰度明显不同（P<0.05），差异丰度miRNA的靶基因富集于不同通路。
在实验条件下，电磁脉冲辐射可改变外泌体的数量和miRNA丰度，进而影响肿瘤来源外泌体的功能。
