巨噬细胞J774A.1外泌体促进结直肠癌细胞系MC38和CT26的增殖和迁移

基础医学与临床 ›› 2021, Vol. 41 ›› Issue (6): 799-804.

巨噬细胞J774A.1外泌体促进结直肠癌细胞系MC38和CT26的增殖和迁移

杨旭东, 于红亮, 李静, 赵春华^*

中国医学科学院基础医学研究所北京协和医学院基础学院细胞生物学系, 北京 100005

收稿日期:2021-03-08 修回日期:2021-04-16 出版日期:2021-06-05 发布日期:2021-05-31
通讯作者: ^*zhaochunhua@vip.163.com
基金资助:
国家自然科学基金(81971324);中国医学科学院医学与健康科技创新工程(2017-I2M-3-007)

Macrophage J774A.1 exosome promotes the proliferation and migration of colorectal cancer cell lines MC38 and CT26

YANG Xu-dong, YU Hong-liang, LI Jing, ZHAO Chun-hua^*

Department of Cell Biology, Institute of Basic Medical Sciences CAMS, School of Basic Medicine PUMC, Beijing 100005, China

Received:2021-03-08 Revised:2021-04-16 Online:2021-06-05 Published:2021-05-31
Contact: ^*zhaochunhua@vip.163.com

摘要/Abstract

摘要： 目的探讨巨噬细胞J774A.1分泌的外泌体对小鼠结直肠癌细胞系MC38和CT26增殖和迁移的影响及作用机制。方法应用流式细胞术对J774A.1细胞进行表型鉴定;应用透射电镜、纳米颗粒跟踪分析(NTA)和Western blot分别检测J774A.1细胞外泌体的形态特征、大小和表面标志物;荧光显微镜下观察MC38和CT26细胞对外泌体的摄取情况;应用CCK8法检测外泌体对MC38和CT26细胞增殖作用的影响;细胞划痕实验检测MC38和CT26细胞的迁移能力;Western blot检测增殖相关通路的磷酸化蛋白p-MEK1/2和p-ERK1/2的表达变化。结果 J774A.1细胞为表达细胞表面标志分子CD68和CD206的M2型巨噬细胞。J774A.1细胞的外泌体均能被结直肠癌细胞MC38和CT26摄取,而且能够促进它们增殖(P＜0.05)、迁移及MEK-ERK信号通路激活。结论巨噬细胞J774A.1的外泌体能够促进结直肠癌细胞系MC38和CT26的增殖及迁移,这可能与MEK-ERK信号通路的磷酸化增加有关。

关键词: 外泌体, 巨噬细胞J774A.1, 结直肠癌细胞, 增殖, 迁移

Abstract: Objective To investigate the effects of exosome secreted by macrophage J774A.1 on the proliferation and migration of mouse colorectal cancer cell lines MC38 and CT26 and potential mechanism. Methods The phenotype of J774A.1 cells was identified by flow cytometry.The morphological characteristics, size and surface markers of the exosome of J774A.1 cells were detected by transmission electronic microscopy, nanoparticle tracking analysis(NTA) and Western blot.The uptake of exosome by MC38 and CT26 cells was observed by fluorescence microscopy. Proliferation of MC38 and CT26 was detected by CCK8 assay. Migration ability of MC38 and CT26 was detected by wound healing assay. The expression of phosphorylated proteins p-MEK1/2 and p-ERK1/2 in proliferation-related pathways was detected by Western blot. Results J774A.1 cells are M2 macrophages expressing the cell surface marker molecules CD68 and CD206. J774A.1 exosomes can be taken up by colorectal cancer cells MC38 and CT26, and can promote their proliferation(P＜0.05), enhance their migration and activate MEK-ERK signaling pathway. Conclusions Macrophage J774A.1 exosomes can promote the proliferation and migration of colorectal cancer cell lines MC38 and CT26, which is potentially attributed to the increased phosphorylation of MEK-ERK signaling pathway.

Key words: exosome, macrophage J774A.1, colorectal cancer cell, proliferation, migration

中图分类号:

R73-37

杨旭东, 于红亮, 李静, 赵春华. 巨噬细胞J774A.1外泌体促进结直肠癌细胞系MC38和CT26的增殖和迁移[J]. 基础医学与临床, 2021, 41(6): 799-804.

YANG Xu-dong, YU Hong-liang, LI Jing, ZHAO Chun-hua. Macrophage J774A.1 exosome promotes the proliferation and migration of colorectal cancer cell lines MC38 and CT26[J]. Basic & Clinical Medicine, 2021, 41(6): 799-804.

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