Kindlin-2 regulates migration and adhesion of vascular smooth muscle cells via β1-integrin
10.3760/cma.j.issn.0253-3758.2014.11.013
- VernacularTitle:Kindlin-2对血管平滑肌细胞迁移和黏附的影响及其机制的实验研究
- Author:
Xiaolin WU
1
;
Hong JIANG
;
Jing CHEN
;
Qi HU
;
Jing ZHANG
;
Jichun WANG
Author Information
1. 430060,武汉大学人民医院心血管内科
- Keywords:
Myocytes,smooth muscle;
Blood vessels;
Cell movement;
Cell adhesion;
Integrins;
Kindlin-2
- From:
Chinese Journal of Cardiology
2014;42(11):938-943
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effects of Kindlin-2 RNA interference on vascular smooth muscle cells (VSMCs) migration,adhesion and β1-integrin as well as the relationship between Kindlin-2 and β1-integrin.Methods Primary VSMCs were cultured,infected with Kindlin-2 siRNA lentiviral vectors.VSMCs were divided into three groups:the blank control group,the negative control group and the Kindlin-2 siRNA group.The ability of VSMCs migration was measured by Transwell experiment and wound healing assay.The ability of VSMCs adhesion to extracelluar matrix was determined by cell-extracelluar matrix adhesion assay.The Kindlin-2 and β1-integrin mRNA and protein expression levels were detected by real-time quantitative PCR and Western blot.Total β1-integrin and active β1-integrin expression on the surface of VSMCs was evaluated by flow cytometry.Results The efficiency of Kindlin-2 siRNA lentivirus infected VSMCs was more than 90％.The number of VSMCs migration in the Kindlin-2 siRNA group was significantly lower than that of the blank control group and the negative group (all P ＜ 0.05).Moreover,the distance of VSMCs migration was shorter than that of the blank control group and the negative group (all P ＜0.05).The number of VSMCs adhesion to collagen Ⅰ was less than that of the blank control group and the negative group (all P ＜ 0.05).A590nm of the Kindlin-2 siRNA group was also lower than that of the blank control group and the negative group (all P ＜ 0.05).Compared with the blank control group,the expression level of Kindlin-2 mRNA in the Kindlin-2 siRNA group decreased 47％ (P ＜ 0.05),but the expression level of β1-integrin mRNA remained unchanged.The Kindlin-2 protein level in the Kindlin-2 siRNA group was lower than that of the blank control group and the negative group (all P ＜ 0.05).β1-integrin protein level was similar among the three groups.Activated β1-integrin on the surface of VSMCs in the Kindlin-2siRNA group was lower than that of the blank control group and the negative group (all P ＜ 0.05).However,the expression level of total β1-integrin on the VSMCs surface was similar among the three groups.Conclusion Kindin-2 can regulate VSMCs migration and adhesion and activate β1-integrin on the surface of VSMCs.
