title: Carboxyl-terminal truncated HBx regulates a distinct microRNA transcription program in hepatocellular carcinoma development.
abstract: BACKGROUND
  The biological pathways and functional properties by which misexpressed microRNAs (miRNAs) contribute to liver carcinogenesis have been intensively investigated. However, little is known about the upstream mechanisms that deregulate miRNA expressions in this process. In hepatocellular carcinoma (HCC), hepatitis B virus (HBV) X protein (HBx), a transcriptional trans-activator, is frequently expressed in truncated form without carboxyl-terminus but its role in miRNA expression and HCC development is unclear.
  
  
  METHODS
  Human non-tumorigenic hepatocytes were infected with lentivirus-expressing full-length and carboxyl-terminal truncated HBx (Ct-HBx) for cell growth assay and miRNA profiling. Chromatin immunoprecipitation microarray was performed to identify the miRNA promoters directly associated with HBx. Direct transcriptional control was verified by luciferase reporter assay. The differential miRNA expressions were further validated in a cohort of HBV-associated HCC tissues using real-time PCR.
  
  
  RESULTS
  Hepatocytes expressing Ct-HBx grew significantly faster than the full-length HBx counterparts. Ct-HBx decreased while full-length HBx increased the expression of a set of miRNAs with growth-suppressive functions. Interestingly, Ct-HBx bound to and inhibited the transcriptional activity of some of these miRNA promoters. Notably, some of the examined repressed-miRNAs (miR-26a, -29c, -146a and -190) were also significantly down-regulated in a subset of HCC tissues with carboxyl-terminal HBx truncation compared to their matching non-tumor tissues, highlighting the clinical relevance of our data.
  
  
  CONCLUSIONS
  Our results suggest that Ct-HBx directly regulates miRNA transcription and in turn promotes hepatocellular proliferation, thus revealing a viral contribution of miRNA deregulation during hepatocarcinogenesis.
authors.affiliation: Institute of Digestive Disease, The Chinese University of Hong Kong, Hong Kong SAR, China.
authors.forename: Wing-Kit
authors.initials: WK
authors.lastname: Yip
authors.forename: Alfred Sze-Lok
authors.initials: AS
authors.lastname: Cheng
authors.forename: Ranxu
authors.initials: R
authors.lastname: Zhu
authors.forename: Raymond Wai-Ming
authors.initials: RW
authors.lastname: Lung
authors.forename: Daisy Pui-Fong
authors.initials: DP
authors.lastname: Tsang
authors.forename: Suki Shuk-Kei
authors.initials: SS
authors.lastname: Lau
authors.forename: Yangchao
authors.initials: Y
authors.lastname: Chen
authors.forename: Jonathan Gabriel
authors.initials: JG
authors.lastname: Sung
authors.forename: Paul Bo-San
authors.initials: PB
authors.lastname: Lai
authors.forename: Enders Kai-On
authors.initials: EK
authors.lastname: Ng
authors.forename: Jun
authors.initials: J
authors.lastname: Yu
authors.forename: Nathalie
authors.initials: N
authors.lastname: Wong
authors.forename: Ka-Fai
authors.initials: KF
authors.lastname: To
authors.forename: Vincent Wai-Sun
authors.initials: VW
authors.lastname: Wong
authors.forename: Joseph Jao-Yiu
authors.initials: JJ
authors.lastname: Sung
authors.forename: Henry Lik-Yuen
authors.initials: HL
authors.lastname: Chan
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000473:pathology*; D002471:Cell Transformation, Neoplastic; D047369:Chromatin Immunoprecipitation; D003001:Cloning, Molecular; D015331:Cohort Studies; D017931:DNA Primers; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000473:pathology*; D035683:MicroRNAs / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D060888:Real-Time Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D015534:Trans-Activators / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D014158:Transcription, Genetic; D054334:Viral Regulatory and Accessory Proteins
chemical_list: D017931:DNA Primers; D035683:MicroRNAs; D015534:Trans-Activators; D054334:Viral Regulatory and Accessory Proteins; C057628:hepatitis B virus X protein