title: [Kindlin-2 regulates migration and adhesion of vascular smooth muscle cells via β1-integrin].
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). A(590 nm) 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-2 siRNA 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.
  
  
  CONCLUSIONS
  Kindin-2 can regulate VSMCs migration and adhesion and activate β1-integrin on the surface of VSMCs.
authors.affiliation: Department of Cardiology, Renmin Hospital of Wuhan University, Wuhan 430060, China.
authors.forename: Xiaolin
authors.initials: X
authors.lastname: Wu
authors.affiliation: Department of Cardiology, Renmin Hospital of Wuhan University, Wuhan 430060, China. Email: jiang_hongw@aliyun.com.
authors.forename: Hong
authors.initials: H
authors.lastname: Jiang
authors.affiliation: Department of Cardiology, Renmin Hospital of Wuhan University, Wuhan 430060, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Chen
authors.affiliation: Department of Cardiology, Renmin Hospital of Wuhan University, Wuhan 430060, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Hu
authors.affiliation: Department of Cardiology, Renmin Hospital of Wuhan University, Wuhan 430060, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Department of Cardiology, Renmin Hospital of Wuhan University, Wuhan 430060, China.
authors.forename: Jichun
authors.initials: J
authors.lastname: Wang
mesh_terms: D015153:Blotting, Western; D002448:Cell Adhesion; D002465:Cell Movement; D002478:Cells, Cultured; D003094:Collagen; D005434:Flow Cytometry; D006801:Humans; D019012:Integrin beta1 / Q000502:physiology*; D008565:Membrane Proteins / Q000502:physiology*; D009131:Muscle, Smooth, Vascular / Q000502:physiology*; D032389:Myocytes, Smooth Muscle; D009363:Neoplasm Proteins / Q000502:physiology*; D034622:RNA Interference; D012333:RNA, Messenger; D034741:RNA, Small Interfering
chemical_list: C477322:FERMT3 protein, human; D019012:Integrin beta1; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D003094:Collagen