title: [Construction and identification of eukaryotic expression plasmid expressing siRNA targeting USP22 gene].
abstract: OBJECTIVE
  To construct eukaryotic expression plasmid expressing siRNA targeting ubiquitin specific peptidase 22 gene (USP22), and to investigate its effect on the growth of hepatoma carcinoma cells HepG2.
  
  
  METHODS
  siRNA templates were synthesized based on USP22 mRNA sequence and cloned into vector Pmscv/Hyg/U6. The resulting recombinant was identified by restriction enzyme digestion and DNA sequencing. Recombinants were than transfected into HepG2 cells mediated by liposome. The USP22 protein and mRNA in HepG2 cells were detected by western blot and RT-PCR, respectively. The cellular growth activity was evaluated with MTT assay.
  
  
  RESULTS
  Recombinant plasmid expressing siRNA targeting USP22 was successfully constructed. The down-regulated protein and mRNA level of USP22 and decreased cellular growth in HepG2 cells transfected with recombinant plasmid were observed.
  
  
  CONCLUSIONS
  The eukaryotic expression vector for RNA interference USP22 gene is constructed successfully, which inhibits the expression of USP22 in HepG2 cells and suppresses cell proliferation.
authors.affiliation: Department of Pharmacology, College of Basic Medical Science, Jiujiang University, Jiujiang 332000, China.
authors.forename: Jian-jun
authors.initials: JJ
authors.lastname: Xiong
authors.forename: Li-jun
authors.initials: LJ
authors.lastname: Gan
authors.forename: Ling
authors.initials: L
authors.lastname: Lin
authors.forename: Zhen
authors.initials: Z
authors.lastname: Gong
authors.forename: Zhou-huan
authors.initials: ZH
authors.lastname: Wu
authors.forename: Wei-dong
authors.initials: WD
authors.lastname: Li
mesh_terms: D001483:Base Sequence; D049109:Cell Proliferation; D005822:Genetic Vectors / Q000235:genetics*; D056945:Hep G2 Cells; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D013869:Thiolester Hydrolases / Q000235:genetics*; D014162:Transfection; D043222:Ubiquitin Thiolesterase
chemical_list: D012333:RNA, Messenger; D034741:RNA, Small Interfering; D013869:Thiolester Hydrolases; D043222:Ubiquitin Thiolesterase; C527170:Usp22 protein, human