title: High load hepatitis B virus replication inhibits hepatocellular carcinoma cell metastasis through regulation of epithelial-mesenchymal transition.
abstract: OBJECTIVE
  The aims of this study were to investigate the effect of hepatitis B virus (HBV) replication on the metastatic ability of hepatocellular carcinoma (HCC) cells and to explore a potential mechanism from the perspective of epithelial-mesenchymal transition (EMT).
  
  
  METHODS
  Two short-interfering RNAs (siRNAs) against the HBV S gene were used to inhibit HBV replication in HepG2.2.15 cells. To evaluate the level of HBV replication and interference efficiency, HBV antigen and HBV DNA were detected by ELISA and quantitative PCR (Q-PCR). Invasion and metastatic abilities were compared between different groups by wound healing and trans-well assays. Immunofluorescent staining and Western blotting were utilized to detect EMT markers.
  
  
  RESULTS
  Both siRNAs effectively inhibited HBV replication in HepG2.2.15 cells. Compared to control HepG2.2.15 cells, cells transfected with the siRNAs showed characteristics of the mesenchymal phenotype and augmented their ability to invade and metastasize. Inhibition of HBV replication suppressed E-cadherin and induced a switch to vimentin expression. Western blots confirmed the decrease in E-cadherin expression. The level of E-cadherin expression was also lower in HepG2 cells than in HepG2.2.15 cells.
  
  
  CONCLUSIONS
  siRNAs were able to effectively inhibit HBV replication in vitro. A high load of HBV replication may inhibit the invasion and metastatic ability of HCC cells by reversing the EMT process.
authors.affiliation: Department of Pathology, Basic Medical Science College, Harbin Medical University, 157 Baojian Road, Nangang District, Harbin 150081, China.
authors.forename: Tianzhen
authors.initials: T
authors.lastname: Wang
authors.affiliation: Department of Pathology, Basic Medical Science College, Harbin Medical University, 157 Baojian Road, Nangang District, Harbin 150081, China.
authors.forename: Yinji
authors.initials: Y
authors.lastname: Jin
authors.affiliation: Department of Pathology, Basic Medical Science College, Harbin Medical University, 157 Baojian Road, Nangang District, Harbin 150081, China.
authors.forename: Ran
authors.initials: R
authors.lastname: Zhao
authors.affiliation: Department of Pathology, Basic Medical Science College, Harbin Medical University, 157 Baojian Road, Nangang District, Harbin 150081, China.
authors.forename: Yiqi
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Department of Pathology, Basic Medical Science College, Harbin Medical University, 157 Baojian Road, Nangang District, Harbin 150081, China.
authors.forename: Yuhua
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Obstetrics and Gynecology, First Affiliated Hospital of Harbin Medical University, Harbin, China.
authors.forename: Di
authors.initials: D
authors.lastname: Wu
authors.affiliation: Department of Gynecology, Third Affiliated Hospital of Harbin Medical University, Harbin, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Kong
authors.affiliation: Department of Pathology, Basic Medical Science College, Harbin Medical University, 157 Baojian Road, Nangang District, Harbin 150081, China; Heilongjiang Provincial Key Laboratory for Infection and Immunity, Harbin Medical University, Harbin, China. Electronic address: wtzpath@163.com.
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Jin
authors.affiliation: Heilongjiang Provincial Key Laboratory for Infection and Immunity, Harbin Medical University, Harbin, China; Department of Microbiology, Basic Medical Science College, Harbin Medical University, 157 Baojian Road, Nangang District, Harbin 150081, China. Electronic address: Fengminzhang@aliyun.com.
authors.forename: Fengmin
authors.initials: F
authors.lastname: Zhang
mesh_terms: D015820:Cadherins / Q000235:genetics / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000821:virology*; D004279:DNA, Viral / Q000235:genetics; D058750:Epithelial-Mesenchymal Transition; D056945:Hep G2 Cells; D006515:Hepatitis B virus / Q000235:genetics* / Q000502:physiology; D006801:Humans; D008113:Liver Neoplasms / Q000821:virology; D034741:RNA, Small Interfering / Q000235:genetics; D014162:Transfection; D014746:Vimentin / Q000235:genetics / Q000378:metabolism; D019562:Viral Load; D014779:Virus Replication
chemical_list: D015820:Cadherins; D004279:DNA, Viral; D034741:RNA, Small Interfering; D014746:Vimentin
keywords: EMT; HBV; HepG2.2.15; Metastasis; siRNA