title: Viral expression and molecular profiling in liver tissue versus microdissected hepatocytes in hepatitis B virus-associated hepatocellular carcinoma.
abstract: BACKGROUND
  The molecular mechanisms whereby hepatitis B virus (HBV) induces hepatocellular carcinoma (HCC) remain elusive. We used genomic and molecular techniques to investigate host-virus interactions by studying multiple areas of the same liver from patients with HCC.
  
  
  METHODS
  We compared the gene signature of whole liver tissue (WLT) versus laser capture-microdissected (LCM) hepatocytes along with the intrahepatic expression of HBV. Gene expression profiling was performed on up to 17 WLT specimens obtained at various distances from the tumor center from individual livers of 11 patients with HCC and on selected LCM samples. HBV markers in liver and serum were determined by real-time polymerase chain reaction (PCR) and confocal immunofluorescence.
  
  
  RESULTS
  Analysis of 5 areas of the liver showed a sharp change in gene expression between the immediate perilesional area and tumor periphery that correlated with a significant decrease in the intrahepatic expression of HB surface antigen (HBsAg). The tumor was characterized by a large preponderance of down-regulated genes, mostly involved in the metabolism of lipids and fatty acids, glucose, amino acids and drugs, with down-regulation of pathways involved in the activation of PXR/RXR and PPARα/RXRα nuclear receptors, comprising PGC-1α and FOXO1, two key regulators critically involved not only in the metabolic functions of the liver but also in the life cycle of HBV, acting as essential transcription factors for viral gene expression. These findings were confirmed by gene expression of microdissected hepatocytes. Moreover, LCM of malignant hepatocytes also revealed up-regulation of unique genes associated with cancer and signaling pathways, including two novel HCC-associated cancer testis antigen genes, NUF2 and TTK.
  
  
  CONCLUSIONS
  Integrated gene expression profiling of whole liver tissue with that of microdissected hepatocytes demonstrated that HBV-associated HCC is characterized by a metabolism switch-off and by a significant reduction in HBsAg. LCM proved to be a critical tool to validate gene signatures associated with HCC and to identify genes that may play a role in hepatocarcinogenesis, opening new perspectives for the discovery of novel diagnostic markers and therapeutic targets.
authors.forename: Marta
authors.initials: M
authors.lastname: Melis
authors.forename: Giacomo
authors.initials: G
authors.lastname: Diaz
authors.forename: David E
authors.initials: DE
authors.lastname: Kleiner
authors.forename: Fausto
authors.initials: F
authors.lastname: Zamboni
authors.forename: Juraj
authors.initials: J
authors.lastname: Kabat
authors.forename: Jinping
authors.initials: J
authors.lastname: Lai
authors.forename: Giulia
authors.initials: G
authors.lastname: Mogavero
authors.forename: Ashley
authors.initials: A
authors.lastname: Tice
authors.forename: Ronald E
authors.initials: RE
authors.lastname: Engle
authors.forename: Steven
authors.initials: S
authors.lastname: Becker
authors.forename: Charles R
authors.initials: CR
authors.lastname: Brown
authors.forename: Jeffrey C
authors.initials: JC
authors.lastname: Hanson
authors.forename: Jaime
authors.initials: J
authors.lastname: Rodriguez-Canales
authors.forename: Michael
authors.initials: M
authors.lastname: Emmert-Buck
authors.forename: Sugantha
authors.initials: S
authors.lastname: Govindarajan
authors.forename: Michael
authors.initials: M
authors.lastname: Kew
authors.affiliation: Hepatic Pathogenesis Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA. pfarci@niaid.nih.gov.
authors.forename: Patrizia
authors.initials: P
authors.lastname: Farci
mesh_terms: D000368:Aged; D006528:Carcinoma, Hepatocellular / Q000235:genetics* / Q000821:virology; D005260:Female; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral; D006509:Hepatitis B / Q000150:complications* / Q000235:genetics; D006515:Hepatitis B virus / Q000235:genetics* / Q000472:pathogenicity; D022781:Hepatocytes / Q000378:metabolism* / Q000821:virology; D054884:Host-Pathogen Interactions / Q000235:genetics; D006801:Humans; D061346:Laser Capture Microdissection; D008099:Liver / Q000378:metabolism* / Q000821:virology; D008113:Liver Neoplasms / Q000235:genetics* / Q000821:virology; D008297:Male; D008875:Middle Aged; D059467:Transcriptome