激活大麻素受体1诱导的单核巨噬细胞J774A.1的迁移依赖RNA结合蛋白HuR

doi:10.3969/j.issn.1006-7795.2016.01.015

首都医科大学学报 ›› 2016, Vol. 37 ›› Issue (1): 76-82.doi: 10.3969/j.issn.1006-7795.2016.01.015

激活大麻素受体1诱导的单核巨噬细胞J774A.1的迁移依赖RNA结合蛋白HuR

赵中新, 常娜, 盖菁菁, 田蕾, 李丽英

首都医科大学基础医学院细胞生物学系‘肝脏保护与再生调节’北京市重点实验室, 北京 100069

收稿日期:2015-07-09 出版日期:2016-02-21 发布日期:2016-02-01
通讯作者: 李丽英 E-mail:liliying@ccmu.edu.cn
基金资助:
国家中医药管理局中医预防医学重点培育学科建设项目[(2012)170]。

Mouse monocyte/macrophage cell line J774A.1 migration induced by the activation of CB1 depends on HuR

Zhao Zhongxin, Chang Na, Ge Jingjing, Tian Lei, Li Liying

Department of Cell Biology, School of Basic Medical Sciences, Municipal Laboratory for Liver Protection and Regulation of Regeneration, Capital Medical University, Beijing 100069, China

Received:2015-07-09 Online:2016-02-21 Published:2016-02-01
Supported by:
This study is supported by TCM Preventive Medicine Major Cultivate Discipline Construction Project of National Traditional Chinese Medicine[(2012)170].

摘要/Abstract

摘要： 目的研究大麻素受体1(cannabinoid receptor 1, CB1)在单核巨噬细胞迁移中的重要作用以及RNA结合蛋白人抗原R(human antigen R, HuR)参与其中的可能机制。方法选用单核巨噬细胞系J774A.1,应用琼脂糖凝胶电泳和免疫荧光染色技术鉴定J774A.1中CB1以及HuR的表达;ACEA和AM281分别为CB1的药理学激动剂和拮抗剂,应用Boyden chamber法检测ACEA和AM281对J774A.1迁移活性的影响。HuR的基因干扰用于确定激活CB1诱导的J774A.1迁移功能是否依赖HuR;胞质蛋白的分离用于探究激活CB1是否能引起J774A.1胞质中HuR的富集;RT-qPCR和Western blotting法检测CB1和HuR mRNA和蛋白质的变化情况。结果该研究证明J774A.1在基因和蛋白质水平上均表达CB1和HuR;激活CB1能够促进J774A.1的迁移(P<0.01)并且能够被其药理学拮抗剂AM281所抑制;激活CB1诱导的J774A.1的迁移依赖HuR;激活CB1促进了J774A.1胞质中HuR的富集进一步影响了CB1的表达,由此HuR参与了激活CB1诱导的J774A.1的迁移。结论激活CB1能够诱导单核巨噬细胞系J774A.1的迁移,且此过程依赖RNA结合蛋白HuR。

关键词: 人抗原R, 大麻素受体1, 细胞迁移

Abstract: Objective To investigate the effect of activation of CB1 on J774A.1 migration and to know whether HuR is involved in the process. Methods Immunofluorescence and agarose gel electrophoresis were used to detect the expression of cannabinoid receptor 1 (CB1) and human antigen R (HuR). Boyden chamber was used for cell migration assay. HuR siRNA transfection was used to determine whether ACEA(CB1 agonist) induced-migration depended on HuR. In order to observe its underlying mechanisms, HuR subcellular localization was assayed by Western blotting of cellular fractions from J774A.1 stimulated with ACEA.The effects of HuR siRNA on CB1 mRNA and protein levels in J774A.1 stimulated with ACEA was assayed by Western blotting. Results In this study, we present evidence that J774A.1 significantly expressed HuR and CB1; the treatment of ACEA caused a increase in J774A.1 migration(P<0.01), and pharmacological inhibition of CB1 markedly attenuated ACEA-induced migration; ACEA-induced J774A.1 migration depended on HuR. Furthermore, we demonstrated ACEA increased HuR cytoplasmic translocation, and HuR regulated the expression of CB1. Conclusion The activation of CB1 induces J774A.1 migration and HuR is involved in the process.

Key words: human antigen R, cannabinoid receptor 1, cell migration

中图分类号:

赵中新, 常娜, 盖菁菁, 田蕾, 李丽英. 激活大麻素受体1诱导的单核巨噬细胞J774A.1的迁移依赖RNA结合蛋白HuR[J]. 首都医科大学学报, 2016, 37(1): 76-82.

Zhao Zhongxin, Chang Na, Ge Jingjing, Tian Lei, Li Liying. Mouse monocyte/macrophage cell line J774A.1 migration induced by the activation of CB1 depends on HuR[J]. Journal of Capital Medical University, 2016, 37(1): 76-82.

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激活大麻素受体1诱导的单核巨噬细胞J774A.1的迁移依赖RNA结合蛋白HuR

Mouse monocyte/macrophage cell line J774A.1 migration induced by the activation of CB1 depends on HuR

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