Expression of Slit3/Robo signal pathway in mouse aortic smooth muscle cell and its impact on proliferation and migration
10.3760/cma.j.issn.0253-3758.2016.06.016
- VernacularTitle:Slit3/Robo信号通路在小鼠主动脉平滑肌细胞的表达及其对细胞增殖和迁移的作用
- Author:
Wei NI
1
;
Tao LIU
;
Haoyu WANG
;
Lihua LIU
;
Guixiu CHEN
Author Information
1. 637000,川北医学院第二临床医学院 南充市中心医院心内科
- Keywords:
Signal transduction;
Myocytes,smooth muscle;
Cell proliferation;
Cell movement
- From:
Chinese Journal of Cardiology
2016;44(6):542-547
- CountryChina
- Language:Chinese
-
Abstract:
Objective To observe the expression of neural axon guidance molecules Slit3 and Robo receptors in mouse aortic smooth muscle cell(MASMC) and investigate the effect of exogenous Slit3 protein on migration and proliferation of MASMC.Methods The primary cultured MASMC were identified by immunofluorescent assay.The expression of Slit3/Robo signal pathway was detected by RT-PCR and immunocytochemical staining.MASMC were divided into 6 groups:the negative control group (DMEM medium containing bovine serum albumin 86 μg/L),Slit3 0μg/L group (DMEM medium without Slit3),Slit3 24 μg/L group (DMEM medium containing Slit3 24 μg/L),Slit3 40 μg/L group (DMEM medium containing Slit3 40 μg/L),Slit3 80 μg/L group (DMEM medium containing Slit3 80 μg/L) and the positive control group (DMEM medium containing platelet derived growth factor 10 μg/L).The effects of exogenous Slit3 on MASMC proliferation and migration were detected by CCK-8 and scratched cells and transwell chambers respectively.Results (1) The mRNA and protein expressions of Slit2,Slit3,Robo1 and Robo4 were detected in MASMC.mRNA level of Slit2 was lower than Slit3 (P ＜ 0.05) and there were no significant difference between mRNA level of Robo1 and Robo4.(2) The mitogenic responses of MASMC were significantly enhanced in Slit3 24 μg/L group,Slit3 40 μg/L group and Slit3 80 μg/L group compared with negative control group (1.13 ± 0.04,1.19 ± 0.02,1.18 ± 0.08 and 0.64 ± 0.10 respectively,all P ＜0.05).The mitogenic activity of MASMC was the strongest in Slit3 40 μg/L group (compared with positive control group 1.27 ± 0.05,P ＞ 0.05).(3) The autonomous migration activity of MASMC were significantly increased in Slit3 24 μg/L group,Slit3 40 μg/L group,Slit3 80 μg/L group compared with negative control group (cell scratch width were (0.40 ± 0.03) cm,(0.32 ± 0.03) cm,(0.30 ± 0.02) cm and (0.49 ±0.01) cm respectively,all P ＜ 0.05).The autonomous migration activity of MASMC was the strongest in Slit3 80 μg/L group (compared with positive control group (0.22 ± 0.01)cm,P＞ 0.05).The transmembrane migration activity of MASMC were significantly increased in Slit3 24 μg/L group,Slit3 40 μg/L group,Slit3 80 μg/L group compared with negative control group (the number of cell migration were 46.67 ± 2.23,65.33 ± 3.43,81.67 ± 4.22 and 39.33 ± 2.03 respectively,all P ＜ 0.05).The transmembrane migration activity of MASMC was the strongest in Slit3 80 μg/L group (compared with positive control group 84.00 ± 2.02,P ＞ 0.05).Conclusion Slit2,Slit3,Robo1 and Robo4 were expressed in MASMC,and exogenous Slit3 could promote proliferation and migration of MASMC in vitro.
