电磁脉冲对肿瘤外泌体介导的非小细胞肺癌转移的干预作用研究

Non-small cell lung cancer with high incidence and mortality is not sensitive to radiotherapy and chemotherapy, lack of ideal treatment methods for now, and tumor metastasis in vivo is the main reason for lacking cure methods and poor prognosis. Lots of researches have confirmed that tumor exosomes play a crucial role in non-small cell lung cancer metastasis in recent years, and they are the potential target of electromagnetic pulse due to their specific electromagnetic properties, so we put forward the hypothesis that electromagnetic pulse can be applied in intervening with non-small cell lung cancer metastasis by targeting tumor exosomes. In this study, at first, by using nanotracer particles analysis and transcriptionomic techniques and Transwell and some other methods, the influence of electromagnetic pulse on the quantity and inclusions of tumor exosomes, the influence of electromagnetic pulse on cell epithelial to mesenchymal transition mediated by them, and some other electromagnetic pulse intervening effects would be explored in vitro, then by using in vivo fluorescence imaging technology and immunohistochemical analysis and some other methods, the influence of electromagnetic pulse on tumor metastasis mediated by them would be explored in vivo. The study aims to clarify the intervening effects of electromagnetic pulse on non-small cell lung cancer metastasis mediated by tumor exosomes, and to put forward effective parameters of electromagnetic pulse, which would provide a novel treatment technology hopefully for non-small cell lung cancer in future.

非小细胞肺癌是一种高发病率和高死亡率癌症，对放化疗均不敏感，至今缺乏理想的治疗手段，肿瘤细胞的体内转移是导致该病无法治愈和患者预后不良的主要原因。最新研究证实，肿瘤外泌体在非小细胞肺癌转移过程中发挥关键作用，并且肿瘤外泌体具有独特的电磁特性，基于此，提出假设：以肿瘤外泌体为靶点应用电磁脉冲可干预非小细胞肺癌转移。本研究首先以非小细胞肺癌细胞和肿瘤外泌体为研究对象，采用纳米颗粒示踪分析、转录组学技术和Transwell等技术手段，从离体水平探讨电磁脉冲对肿瘤外泌体数量、内含物和介导肿瘤细胞上皮-间质转化等的影响，然后以肺癌动物模型为研究对象，采用在体荧光成像技术和免疫组化分析等技术手段，从在体水平探讨电磁脉冲对肿瘤外泌体介导的肿瘤转移的影响。本研究旨在明确电磁脉冲对肿瘤外泌体介导的非小细胞肺癌转移的干预作用，提出电磁脉冲有效作用参数，为未来非小细胞肺癌的治疗提供新的技术手段。

大量研究证实，肿瘤外泌体（TDEs）在非小细胞肺癌发展和转移过程中扮演者关键角色，而电磁脉冲（EMP）可通过多种机制延缓肿瘤的发展，但EMP与TDEs的关系尚未研究。该研究以非小细胞肺癌细胞A549为主要研究对象，结合在体荷瘤模型，一是开展了EMP对A549细胞分泌TDEs数量和外泌体内miRNA的影响，纳米颗粒追踪分析（NTA）和RNA-seq结果显示，不同强度的EMP可明显影响A549细胞分泌TDEs的数量和TDEs内含miRNA的丰度；二是开展了EMP和外泌体生成抑制剂GW4869对A549细胞恶性表型的影响和EMP对巨噬细胞吸收TDEs及功能的影响，研究发现，EMP和GW4869单独或联合应用不影响A549细胞的增殖、细胞周期分布、细胞凋亡和上皮-间质转化，EMP也不影响巨噬细胞吸收TDEs和分泌TNF-a，但EMP和GW4869联合应用可明显抑制A549细胞的迁移能力；三是开展了EMP和GW4869对体内非小细胞肺癌生长发展的研究，研究发现，GW4869单独以及和EMP联合应用，不影响体内瘤块的体积、重量、组织形态和ki67阳性细胞的数量。该研究为明确EMP与TDEs的关系以及EMP在肿瘤治疗方面的临床应用提供了参考。