title: [Effect of Knockdown of Vascular Cell Adhesion Molecule-1 on the Immunologic Regulation Capacity of Murine Mesenchymal Stem Cells].
abstract: OBJECTIVE
  To establish the stably lower expression of vascular cell adhesion molecule-1 (VCAM-1) in MSC cell line (C3H10T1/2) by siRNA technology, and explore the effect of knockdown of VCAM-1 on the immunologic regulation capacity of murine MSC.
  
  
  METHODS
  The mouse GV118-VCAM-1-RNAi retrovirus vector was constructed by gene recombination technology. The recombinant plasmid was identified by restriction analysis and sequencing, and then the recombinant plasmid GV118-VCAM-1-RNAi was transfected into 293 cells by Lipofectamine, and the supernatant was collected to transfect C3H10T1/2. Moreover, the VCAM-1 lower expression on MSC was evaluated by flow cytometry and fluorescent microscopy. The knockdown VCAM-1 MSC was sorted by flow cytometry. Furthermore, the inhibitory effect of the knockdown VCAM-1 MSC on lymphocyte proliferation was tested by lymphoblast transformation assay (LTT) and mixed lymphocyte reaction assay(MLR).
  
  
  RESULTS
  The recombinant retroviral vector of knockdown VCAM-1 (GV118-VCAM-1-RNAi) was successfully constructed and transfected into mouse MSC cell line C3H10T1/2. The knockdown VCAM-1/MSC was obtained by flow cytometric sorting. The LTT and MLR assay showed that the immunosuppressive effect of MSC lower-expressing VCAM-1 dramatically decreased (P<0.05).
  
  
  CONCLUSIONS
  Knockdown VCAM-1 in MSC can significantly down-regulate the inhibitory capability of MSC on the proliferation of T-cells. The data of this study laid an experimental foundation for studying effect of VCAM-1 transfecting into MSC on immune function.
authors.affiliation: Department of Pediatrics, General Hospital of Tianjin Medical University, Tianjin 30052, China.|Department of Cell Biology, Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850.
authors.forename: Yong-Qing
authors.initials: YQ
authors.lastname: Ni
authors.affiliation: Department of Cell Biology, Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850.
authors.forename: Yuan-Lin
authors.initials: YL
authors.lastname: Liu
authors.affiliation: Department of Cell Biology, Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850.
authors.forename: Heng
authors.initials: H
authors.lastname: Zhu
authors.affiliation: Department of Pediatrics, General Hospital of Tianjin Medical University, Tianjin 30052, China.
authors.forename: Li-Hui
authors.initials: LH
authors.lastname: Wang
authors.affiliation: Department of Pediatrics, General Hospital of Tianjin Medical University, Tianjin 30052, China.
authors.forename: Shi-Feng
authors.initials: SF
authors.lastname: Ma
authors.affiliation: Department of Postgraduate, Hebei North College, Zhangjiakou 07500, Heibei Province, China.
authors.forename: Xiu-Hui
authors.initials: XH
authors.lastname: Chen
authors.affiliation: Department of Pediatrics, General Hospital of Tianjin Medical University, Tianjin 30052, China. E-mail: rxzheng@hotmail.com.
authors.forename: Rong-Xiu
authors.initials: RX
authors.lastname: Zheng
authors.affiliation: Department of Cell Biology, Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850. E-mail: zhangyi612@hotmail.com.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000818:Animals; D002460:Cell Line; D002465:Cell Movement; D049109:Cell Proliferation; D005434:Flow Cytometry; D005822:Genetic Vectors; D008213:Lymphocyte Activation; D059630:Mesenchymal Stem Cells; D051379:Mice; D010957:Plasmids; D034622:RNA Interference; D034741:RNA, Small Interfering; D013601:T-Lymphocytes; D014162:Transfection; D019010:Vascular Cell Adhesion Molecule-1
chemical_list: D034741:RNA, Small Interfering; D019010:Vascular Cell Adhesion Molecule-1