title: [Inhibition of HBsAg and HBeAg secretion by RNA interference of the polymerase gene sequence of hepatitis B virus: an experimental study].
abstract: OBJECTIVE
  To investigate the effects of small interfering RNA (siRNA) targeting the polymerase (P) gene sequence of hepatitis B virus (HBV) on the replication and antigen secretion of HBV.
  
  
  METHODS
  From the 29 base sequences of the HBV in the HepG2.2.15 cells that accord with the demands of siRNA designing five sequences targeting the P gene of HBV were selected and cloned into the siRNA expressing vector pGE-1. Then the plasmid pGE-HBVP was transfected into the cultured HepG2.2.15 cells. Chemiluminescent immunoassay was used to determine the levels of HBsAg and HBeAg in the supernatant of culture medium 24, 48, 72, and 96 hours after the transfection and the expression of HBsAg in the 2.2.15 cells 24 hours after the transfection so as to observe the inhibitory effects. Untransfected cells and cells transfected with blank pGE-1 vector were used as controls.
  
  
  RESULTS
  Five vectors expressing the siRNAs targeting the HBV P region, pGE-HBVP1-pGE-HBV5 were successfully constructed. The efficiency of transfection of the vectors into the 2.2.15 cells were 30% to 40%. 24, 48, 72, and 96 hours after the transfection of pGE-HBVP2, the strongest inhibitor among the five, the inhibitory rates of HBsAg secretion in the supernatant were 28.88%, 32.28%, 29.10%, and 18.42% respectively; and the inhibitory rates of HBeAg secretion were 38.33%, 27.50%, 33.41%, and 12.60% respectively. In view of the transfection efficiency of 30%-40%, the actual inhibitory rate of HBV antigen secretion might reach 80% and over. 24 hours after the transfection the expression rate of HBsAg in the 2.2.15 cells transfected with pGE-HBVP2 was 50%, significantly lower than that in the cells transfected with the blank vector pGE-1 (82%).
  
  
  CONCLUSIONS
  siRNAs targeting the HBV P gene effectively prevent the HBV gene expression and replication and may play an important role in the clinical anti-viral treatment.
authors.affiliation: Beijing Ditan Hospital, Beijing 100011, China.
authors.forename: Shun-ai
authors.initials: SA
authors.lastname: Liu
authors.forename: Hong-shan
authors.initials: HS
authors.lastname: Wei
authors.forename: Qing-ming
authors.initials: QM
authors.lastname: Dong
authors.forename: Jing-jing
authors.initials: JJ
authors.lastname: Guo
authors.forename: Jing
authors.initials: J
authors.lastname: Qin
authors.forename: Qian-ying
authors.initials: QY
authors.lastname: Zhang
authors.forename: Zhi-ying
authors.initials: ZY
authors.lastname: Liu
authors.forename: Jie
authors.initials: J
authors.lastname: Yan
authors.forename: Jun
authors.initials: J
authors.lastname: Cheng
mesh_terms: D003086:Bacteriocin Plasmids / Q000235:genetics; D045744:Cell Line, Tumor; D015684:Gene Products, pol / Q000235:genetics / Q000378:metabolism*; D006514:Hepatitis B Surface Antigens / Q000378:metabolism*; D006513:Hepatitis B e Antigens / Q000378:metabolism*; D006515:Hepatitis B virus / Q000201:enzymology / Q000235:genetics / Q000276:immunology; D006801:Humans; D007118:Immunoassay; D034622:RNA Interference; D014162:Transfection
chemical_list: D015684:Gene Products, pol; D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens; C066027:P protein, Hepatitis B virus