摘要
外泌体(exosome)是直径约30～150 nm的由细胞分泌的一种具有生物学活性的囊泡。有些来自癌细胞的外泌体可以将巨噬细胞(macrophages,Mφ)极化为M2亚型,但前列腺癌细胞来源的外泌体在巨噬细胞极化中的作用仍缺乏研究。本研究采用超滤法提取前列腺癌细胞PC-3M-2B4和PC-3M-IE8条件培养基中的外泌体(PCa-exo)。分别用透射电子显微镜、纳米粒径分析及Western印迹对外泌体形态、颗粒大小和表面的特异性分子标志进行分析鉴定。用PKH67标记外泌体,观察PCa-exo能否被巨噬细胞吸收。免疫荧光分析PCa-exo处理巨噬细胞后,M2型巨噬细胞表面分子标志CD206的表达差异。用q-PCR观察PCa-exo诱导后的巨噬细胞中IL-10、IL-1β等细胞因子的表达。电镜、Western印迹和纳米粒径分析的结果显示,PCa-exo形态多为圆形,直径约为40～150 nm,PCa-exo能被巨噬细胞大量吸收。PCa-exo诱导后,巨噬细胞中CD206荧光表达显著增高,IL-10、IL-1β及IL-12等炎症因子的表达水平与M2/TAM亚型巨噬细胞的表达谱一致。本研究表明,前列腺癌细胞来源的外泌体能诱导巨噬细胞极化为M2表型。
Exosomes are biologically active vesicles secreted by cells with a diameter of 30-150 nm.Some exosomes from cancer cells can polarize macrophages( Mφ) into M2 subtypes. However,the role of prostate cancer-derived exosomes in macrophage polarization remains unknown. In this study,ultrafiltration was used to extract exosomes( PCa-exo) from conditioned medium of prostate cancer cells PC-3 M-2 B4 and PC-3 M-IE8. Transmission electron microscopy,nanoparticle size analysis and Western blotting were used to analyze and identify the shape,particle size and specific surface molecular markers of exosomes respectively. Exosomes were labeled with PKH67 to observe whether PCa-exo could beabsorbed by macrophages. The differential expression of CD206 on M2 macrophages was detected by immunofluorescence. The expression of cytokines such as IL-10 and IL-1β in macrophages induced by PCa-exo was observed by q-PCR. Our results showed that the PCa-exo morphology was mostly round and the diameter was about 40-150 nm. PCa-exo could be absorbed by macrophages in large quantities. After PCa-exo induction,the expression of CD206 in macrophages was significantly increased,and the expression levels of inflammatory factors such as IL-10,IL-1β and IL-12 were consistent with those of M2/TAM subtype macrophages. This study demonstrates that exosome from prostate cancer cells can induce macrophage polarization to the M2 phenotype.
作者
彭一琳
倪煦东
曹帆
刘琦
胡银英
黄艳琴
闵卫平
王志刚
PENG Yi-Lin;Ni Xu-Dong;CAO Fan;LIU Qi;HU Yin-Ying;HUANG Yan-Qin;MIN Wei-Ping;WANG Zhi-Gang(Institute of Cancer Research,Jiangxi Academy of Medical Sciences Nanchang 330006,China;Department of Biochemistry and Molecular Biology,School of Basic Medical Sciences,Nanchang University,Nanchang 330006,China;Institute of Immunology and Biotherapy ,Nanchang University,Nanchang 330006,China)
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2019年第2期179-186,共8页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金(No.81560465)
江西省自然科学基金项目(No.20161BAB205230)资助~~
关键词
前列腺癌细胞
外泌体
巨噬细胞极化
prostate cancer cells
exosomes
macrophage polarization
