Effect of CXCR4 silencing on hypoxia-induced migration of N9 microglia cells
10.3760/cma.j.issn.1671-8925.2014.11.007
- VernacularTitle:CXCR4沉默对缺氧诱导的N9小胶质细胞迁移能力的影响
- Author:
Li CHONG
1
;
Peng TANG
;
Peng LIU
;
Li CHEN
;
Yue LIU
;
Rui LI
;
Minxia GUO
Author Information
1. 陕西省人民医院神经内科
- Keywords:
Microglia;
Hypoxia;
CXCR4;
Gene silencing;
Hypoxia inducible factor-1α
- From:
Chinese Journal of Neuromedicine
2014;13(11):1112-1116
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effect ofhypoxia on CXCR4 expression in N9 microglia cells and the role of CXCR4 in hypoxia-induced N9 cell migration.Methods N9 microglia cells were cultured in normoxia and hypoxia conditions; total cell RNA and protein were prepared,real-time PCR was used to detect the CXCR4 mRNA expression,and Western blotting was employed to detect the protein expressions ofhypoxia-inducible factor-1α (HIF-1α) and CXCR4.After cells were transfected with siRNA targeting HIF-1α or CXCR4,the CXCR4 expression was detected again.The migration ability of N9 cells in normoxia and hypoxia conditions were detected by Transwell assay.After cells were treated with CXCR4 siRNA,the migration ability of N9 cell in hypoxia was measured again.Results As compared with cell cultured in normoxia condition,the CXCR4 expression was significantly increased as compared with cells cultured in hypoxia condition (P＜0.05); however,the hypoxia-induced CXCR4 up-regulation was prevented by HIF-1α silencing.The migration ability of N9 cells in hypoxia condition was significantly higher than that of cells cultured in normoxia condition (number of transmembrane cells:(63.00±5.57) and (20.33±2.08),respectively,P＜0.05); however,the N9 cell migration ability promoted by hypoxia was obviously inhibited by CXCR4 silencing (number of transmembrane cells:63.00±4.00 in si-Control cells and 19.33±3.21 in CXCR4 siRNA cells,P＜0.05).Conclusion Hypoxia can promote the migration ability of N9 cells through inducing CXCR4 expression.
