M2型巨噬细胞来源外泌体对乳腺癌细胞系4T1干性特征的影响

基础医学与临床 ›› 2019, Vol. 39 ›› Issue (6): 798-804.

M2型巨噬细胞来源外泌体对乳腺癌细胞系4T1干性特征的影响

张晨¹,郭建²,赫慧文¹,王胜男¹,陈惠琳¹,段昭君¹,罗云萍¹

1. 中国医学科学院基础医学研究所
2. 中国医学科学院基础医学研究所

收稿日期:2019-03-25 修回日期:2019-04-16 出版日期:2019-06-05 发布日期:2019-06-04
通讯作者: 罗云萍 E-mail:ypluo@ibms.pumc.edu.cn
基金资助:
国家自然科学基金

Effect of M2 macrophage-derived exosomes on stemness of breast cancer cell line 4T1

Received:2019-03-25 Revised:2019-04-16 Online:2019-06-05 Published:2019-06-04
Supported by:
National Natural Science Foundation of China

摘要/Abstract

摘要： 目的探讨M2型巨噬细胞来源的外泌体对乳腺癌细胞系4T1细胞肿瘤干性特征的影响。方法用超速离心法分离M2型巨噬细胞来源的外泌体；用透射电子显微镜、纳米粒子追踪技术，Western blot等对其进行鉴定；然后以乳腺癌细胞系4T1为肿瘤模型，使用Dil染料标记M2型巨噬细胞来源的外泌体，通过免疫荧光技术观察其能否进入4T1细胞。此外，用流式细胞计量术、共培养体系和Transwell实验探究在M2型巨噬细胞来源的外泌体影响下的4T1细胞的干性特征。结果 M2型巨噬细胞在体外被成功诱导，分离并鉴定了M2型巨噬细胞来源的外泌体。通过免疫荧光，看到Dil标记的外泌体可成功进入4T1细胞。M2型巨噬细胞来源的外泌体与4T1细胞共孵育后，可明显增强4T1细胞的迁移能力（P<0.001）和成球能力（P<0.05）；外泌体抑制剂GW4869可明显抑制4T1细胞的成球能力（P<0.05）。同时，M2型巨噬细胞来源的外泌体可明显增加4T1细胞中CD44highCD24low细胞的比例（P<0.001）。结论 M2型巨噬细胞来源的外泌体能够促进4T1肿瘤细胞的迁移和成球能力，并明显增加4T1细胞中肿瘤干细胞的比例。

关键词: M2型巨噬细胞, 外泌体, 细胞迁移, 细胞成球, 肿瘤干细胞

Abstract: Objective To explore the effect of M2 macrophage-derived exosomes on stemness of 4T1 cells. Method M2 macrophage-derived exosomes were isolated by ultracentrifugation. And transmission electron microscopy, nanoparticle tracking technology and Western blot were performed to identified the exosomes. Then, the M2 macrophage-derived exosomes were labeled with Dil which makes it available for us to observe their transfer to breast cancer cell line 4T1 by immunofluorescence. After that, flow cytometry, co-culture system and transwell chamber were used to investigate the tumor phenotype and stemness of 4T1 under the influence of M2 macrophage-derived exosomes. Results First, the M2 macrophages were successfully induced in vitro. In addition, the M2 macrophage-derived exosomes were successfully isolated and verified. M2 macrophage-derived exosomes labeled with Dil could be transferred into 4T1 cells. After co-incubation with M2 macrophage-derived exosomes, the migration (P<0.001) and sphere formation (P<0.05) ability of 4T1 cells were significantly enhanced. While after using exosome inhibitor GW4869, the number and volume of spheres formed by 4T1 stem cells were reduced (P<0.05). The proportion of CD44highCD24low cells in 4T1 cells (P<0.001) was significantly increased after incubating with M2 macrophage-derived exosomes. Conclusions M2 macrophage-derived exosomes can increase the migration and sphere formation ability of 4T1 tumor cells, so is the proportion of tumor stem cells in 4T1 cells.

Key words: M2 macrophage, exosome, migration, sphere formation, tumor stem cell

中图分类号:

R737.9

张晨郭建赫慧文王胜男陈惠琳段昭君罗云萍. M2型巨噬细胞来源外泌体对乳腺癌细胞系4T1干性特征的影响[J]. 基础医学与临床, 2019, 39(6): 798-804.

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