M1型巨噬细胞衍生纳米囊泡介导三阴性乳腺癌光热-免疫治疗的研究  

Photothermal Immunotherapy of Triple Negative Breast Cancer Mediated by M1 Macrophage Derived Nanoparticles

作　　者：郭彦敏 刘杰[2] 李玮[2] 王鸿玲[3] GUO Yan-Min;LIU Jie;LI Wei;WANG Hong-Ling(Department of Immunology,Binzhou Medical University,Yantai 264003,China;Department of Medical Laboratory,Yantai Yuhuangding Hospital,Yantai 264000,China;Department of Material Supply,Yantai Yuhuangding Hospital,Yantai 264000,China)

机构地区：[1]滨州医学院免疫学教研室,烟台264003 [2]烟台毓璜顶医院医学检验科,烟台264000 [3]烟台毓璜顶医院物资材料供应科,烟台264000

出　　处：《分析化学》2025年第3期429-440,共12页Chinese Journal of Analytical Chemistry

基　　金：烟台市科技计划项目(No.2022MSGY074)资助。

摘　　要：光热治疗作为一种新型的抗肿瘤治疗策略,单独用于抗肿瘤治疗时靶向性不足,穿透能力有限,无法实现长期抑制。本研究将M1型巨噬细胞外泌体(MEVs)修饰于搭载有CSF1R-siRNA和光热剂的载体表面制备多功能基因递送载体MEVs@RNPs,可诱导巨噬细胞由M2型重编程为抗肿瘤的M1型,实现三阴性乳腺癌(TNBC)的高效光热-免疫治疗。采用纳米粒度仪、紫外-可见分光光度计、荧光分光光度计和测温热成像仪对纳米颗粒的理化性质进行测定。以TNBC细胞4T1、内皮细胞Bend3和巨噬细胞RAW264.7为体外实验细胞模型,考察了MEVs@RNPs体外靶向肿瘤细胞、杀伤肿瘤细胞和诱导巨噬细胞重编程的能力。选择Balb/c小鼠建立皮下移植瘤模型,测定不同处理组肿瘤体积和体重的变化。分离小鼠血液和肿瘤组织,采用苏木素-伊红(Hematoxylin-Eosin(HE))染色、原位末端转移酶标记(Terminal deoxynucleotidyl transferase dUTP nick end labeling,TUNEL)、酶联免疫吸附分析(Enzyme-linked immunosorbent assay,ELISA)法测定肿瘤细胞的凋亡和细胞因子分泌情况。结果表明,MEVs@RNPs可优先靶向至肿瘤细胞,并在光热条件下诱导更强的TNBC细胞杀伤效果。流式细胞术和ELISA法测定结果显示,MEVs@RNPs可以促进巨噬细胞由M2型向抗肿瘤的M1型转化,并显著提高抗肿瘤细胞因子肿瘤坏死因子-α(TNF-α)和白细胞介素-12(IL-12)的分泌水平。MEVs@RNPs的体内抗肿瘤治疗效果评估结果显示,在激光照射条件下,MEVs@RNPs组可以显著诱导肿瘤细胞病理性坏死和凋亡,促进巨噬细胞重编程和CD4^(+)/CD8^(+)T细胞向肿瘤部位浸润,提高TNF-α和IL-12的分泌水平,从而全面提高TNBC的治疗效果。Photothermal therapy,as a novel anti-tumor treatment strategy,faces many challenges such as insufficient targeting,limited penetration ability,and inability to achieve long-term inhibition when used alone for anti-tumor treatment.Based on this,in this study,macrophage extracellular vesicles(MEVs)were modified onto the surface of a vector carrying CSF1R-siRNA and photothermal agents to form a multifunctional gene delivery vector [e-mail] could induce the macrophages to reprogram from M2 type to M1 type,so as to achieve efficient photothermal and immunotherapy for triple negative breast cancer(TNBC).The basic physicochemical properties of each nanocarrier were characterized using nanoparticle size analyzer,UV-vis spectrometer,fluorescence spectrometer,thermal imaging instrument,etc.TNBC cells 4T1,endothelial cells Bend3,and macrophages 264.7 were used as in vitro experimental subjects for measurement of [e-mail] ability to target tumor cells,kill tumor cells,and induce macrophage reprogramming in vitro was investigated.With Balb/c mice as the in vivo research object,a subcutaneous transplant tumor model was established in mice,and the changes in tumor volume and body weight of mice in different treatment groups were measured.In addition,further isolation of mouse blood and tumor tissue was performed,and hematoxylin-eosin(HE)staining,terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL)method,and enzyme-linked immunosorbent assay(ELISA)method were used to determine the apoptosis and cytokine secretion of tumor cells for comprehensive evaluation of [e-mail] results showed that the MEVs@RNPs could preferentially target tumor cells and induce stronger TNBC cell killing effect under photothermal conditions.Meanwhile,the results of flow cytometry and ELISA analysis showed that the MEVs@RNPs could promote the transformation of macrophages from M2 type to anti-tumor M1 type,significantly enhancing the secretion levels of anti-tumor cytokines tumor necrosis factor-α(TNF-α)and interleukin-12(IL-

关 键 词：光热治疗 免疫治疗 外泌体 巨噬细胞 重编程 三阴性乳腺癌 

分 类 号：R73[医药卫生—肿瘤]