title: Inhibition of hepatitis B virus expression and replication by RNA interference in HepG2.2.15.
abstract: OBJECTIVE
  To observe the inhibition of hepatitis B virus replication and expression by transfecting vector-based small interference RNA (siRNA) pGenesil-HBV X targeting HBV X gene region into HepG2.2.15 cells.
  
  
  METHODS
  pGenesil-HBV X was constructed and transfected into HepG2.2.15 cells via lipofection. HBV antigen secretion was determined 24, 48, and 72 h after transfection by time-resolved immunofluorometric assays (TRFIA). HBV replication was examined by fluorescence quantitative PCR, and the expression of cytoplasmic viral proteins was determined by immunohistochemistry.
  
  
  RESULTS
  The secretion of HBsAg and HBeAg into the supernatant was found to be inhibited by 28.5% and 32.2% (P < 0.01), and by 38.67% (P < 0.05) and 42.86% (P < 0.01) at 48 h and 72 h after pGenesil-HBV X transfection, respectively. Immunohistochemical staining for cytoplasmic HBsAg showed a similar decline in HepG2.2.15 cells 48 h after transfection. The number of HBV genomes within culture supernatants was also significantly decreased 48 h and 72 h post-transfection as quantified by fluorescence PCR (P < 0.05).
  
  
  CONCLUSIONS
  In HepG2.2.15 cells, HBV replication and expression is inhibited by vector-based siRNA pGenesil-HBV X targeting the HBV X coding region.
authors.affiliation: Institute of Hepatology, Changzhi Medical College, Changzhi 046000, Shanxi Province, China. zhaozf_1226@163.com
authors.forename: Zhong-Fu
authors.initials: ZF
authors.lastname: Zhao
authors.forename: Hui
authors.initials: H
authors.lastname: Yang
authors.forename: De-Wu
authors.initials: DW
authors.lastname: Han
authors.forename: Long-Feng
authors.initials: LF
authors.lastname: Zhao
authors.forename: Guo-Ying
authors.initials: GY
authors.lastname: Zhang
authors.forename: Yun
authors.initials: Y
authors.lastname: Zhang
authors.forename: Ming-She
authors.initials: MS
authors.lastname: Liu
mesh_terms: D045744:Cell Line, Tumor; D004261:DNA Replication / Q000235:genetics* / Q000502:physiology; D004279:DNA, Viral / Q000235:genetics; D005786:Gene Expression Regulation / Q000235:genetics / Q000502:physiology; D015967:Gene Expression Regulation, Viral / Q000235:genetics / Q000502:physiology; D005822:Genetic Vectors; D006511:Hepatitis B Antigens / Q000235:genetics / Q000378:metabolism; D006514:Hepatitis B Surface Antigens / Q000235:genetics / Q000378:metabolism; D006513:Hepatitis B e Antigens / Q000235:genetics / Q000378:metabolism; D006515:Hepatitis B virus / Q000235:genetics* / Q000378:metabolism; D018197:Hepatoblastoma / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D010957:Plasmids / Q000235:genetics; D034622:RNA Interference / Q000502:physiology*; D034741:RNA, Small Interfering / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D054334:Viral Regulatory and Accessory Proteins
chemical_list: D004279:DNA, Viral; D006511:Hepatitis B Antigens; D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens; D034741:RNA, Small Interfering; D012367:RNA, Viral; D015534:Trans-Activators; D054334:Viral Regulatory and Accessory Proteins; C057628:hepatitis B virus X protein