title: Effect of vector-expressed siRNA on HBV replication in hepatoblastoma cells.
abstract: OBJECTIVE
  To study the effect of siRNA expressed from DNA vector on HBV replication.
  
  
  METHODS
  Human U6 promoter was amplified from genomic DNA and cloned into plasmid pUC18 to construct a mammalian siRNA expression vector pUC18U6. Then oligonucleotides coding for a short hairpin RNA against HBV were cloned into pUC18U6 to form pUC18U6HBVsir which was introduced into 2.2.15 cells by using liposome-mediated transfection. 2.2.15 cells transfected by pUC18U6 and pUC18U6GFPsir which expressed siRNA against green fluorescent protein and mock-transfected 2.2.15 cells were used as controls. Concentration of HBsAg in the supernatant of the transfected cells was measured by using solid-phase radioimmunoassay.
  
  
  RESULTS
  A mammalian siRNA expression vector pUC18U6 was constructed successfully. Compared with controls, pUC18U6HBVsir which expressed siRNA against HBV decreased concentration of HBsAg significantly by 44%(P<0.05).
  
  
  CONCLUSIONS
  HBV replication in 2.2.15 cells is inhibited by siRNA expressed from the DNA vector.
authors.affiliation: Department of Etiology, Fourth Military Medical University, Xi'an 710033, Shaanxi Province, China. etiology@fmmu.edu.cn
authors.forename: Jun
authors.initials: J
authors.lastname: Liu
authors.forename: Ying
authors.initials: Y
authors.lastname: Guo
authors.forename: Cai-Fang
authors.initials: CF
authors.lastname: Xue
authors.forename: Ying-Hui
authors.initials: YH
authors.lastname: Li
authors.forename: Yu-Xiao
authors.initials: YX
authors.lastname: Huang
authors.forename: Jin
authors.initials: J
authors.lastname: Ding
authors.forename: Wei-Dong
authors.initials: WD
authors.lastname: Gong
authors.forename: Ya
authors.initials: Y
authors.lastname: Zhao
mesh_terms: D045744:Cell Line, Tumor; D005822:Genetic Vectors / Q000235:genetics*; D006509:Hepatitis B / Q000503:physiopathology / Q000821:virology*; D006515:Hepatitis B virus / Q000235:genetics / Q000254:growth & development*; D018197:Hepatoblastoma; D006801:Humans; D010957:Plasmids / Q000235:genetics*; D011401:Promoter Regions, Genetic; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection; D014779:Virus Replication / Q000235:genetics
chemical_list: D034741:RNA, Small Interfering