title: [Construction of eukaryotic expression plasmid expressing siRNA targeting HLA-G gene and detection of its specific downregulation effect].
abstract: OBJECTIVE
  To investigate the effect of HLA-G siRNA on the protection of trophoblast cells from NK cell lysis, an HLA-G-targeting siRNA (small interfering)-expressing plasmid was constructed.
  
  
  METHODS
  HLA-G siRNA plasmid, pSuppressor-U6-neo-HLA-G, was constructed and transfected to JEG-3 cells. The level of messenger RNA and protein of HLA-G in the transfected cells was detected by RT-PCR and Western blot. NK-92MI cells were co-cultured with pSuppressor-U6-neo-HLA-G transfected JEG-3 cells. The cytotoxicity of NK-92MI cells against the transfected cells was analyzed by LDH releasing assay.
  
  
  RESULTS
  Downregulation of HLA-G in HLA-G siRNA plasmid transfected JEG-3 cells was confirmed by RT-PCR and Western blot. Compared with JEG-3 cells without HLA-G without HLA-G siRNA plasmid transfection, the transfected JEG-3 cells showed higher lytic activity.
  
  
  CONCLUSIONS
  HLA-G siRNA can increasing NK cytolysis against JEG-3 cells. HLA-G plays a role in preventing of trophoblast cells from NK cytotoxicity.
authors.affiliation: Department of Obstetrics and Gynecology, Tangdu Hospital, Xi'an 710038, China. jhch0904@163.com
authors.forename: Jian-hui
authors.initials: JH
authors.lastname: Chen
authors.forename: Yuan-qing
authors.initials: YQ
authors.lastname: Yao
authors.forename: Kai-bo
authors.initials: KB
authors.lastname: Hou
authors.forename: Ying-feng
authors.initials: YF
authors.lastname: Lei
authors.forename: Wen
authors.initials: W
authors.lastname: Yin
mesh_terms: D015153:Blotting, Western; D002460:Cell Line; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D015536:Down-Regulation; D006680:HLA Antigens / Q000235:genetics* / Q000378:metabolism; D059951:HLA-G Antigens; D015395:Histocompatibility Antigens Class I / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007694:Killer Cells, Natural / Q000166:cytology / Q000502:physiology; D010957:Plasmids / Q000235:genetics*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D014327:Trophoblasts / Q000166:cytology
chemical_list: D006680:HLA Antigens; D059951:HLA-G Antigens; D015395:Histocompatibility Antigens Class I; D034741:RNA, Small Interfering