title: Analysis of hepcidin expression: in situ hybridization and quantitative polymerase chain reaction from paraffin sections.
abstract: OBJECTIVE
  To establish methods for quantitative polymerase chain reaction (PCR) for hepcidin using RNAs isolated from paraffin-embedded sections and in situ hybridization of hepatocellular carcinoma (HCC).
  
  
  METHODS
  Total RNA from paraffin-embedded sections was isolated from 68 paraffin-embedded samples of HCC. Samples came from 54 male and 14 female patients with a mean age of 66.8 ± 7.8 years. Quantitative PCR was performed. Immunohistochemistry and in situ hybridization for hepcidin were also performed.
  
  
  RESULTS
  Quantitative PCR for hepcidin using RNAs isolated from paraffin-embedded sections of HCC was performed successfully. The expression level of hepcidin mRNA in cancer tissues was significantly higher than that in non-cancer tissues. A method of in situ hybridization for hepcidin was established successfully, and this demonstrated that hepcidin mRNA was expressed in non-cancerous tissue but absent in cancerous tissue.
  
  
  CONCLUSIONS
  We have established novel methods for quantitative PCR for hepcidin using RNAs isolated from paraffin-embedded sections and in situ hybridization of HCC.
authors.affiliation: Second Department of Surgery, School of Medicine, Dokkyo Medical University, Kitakobayashi 880, Mibu, Shimotsuga, Tochigi 321-0293, Japan.
authors.forename: Yuhki
authors.initials: Y
authors.lastname: Sakuraoka
authors.forename: Tokihiko
authors.initials: T
authors.lastname: Sawada
authors.forename: Takayuki
authors.initials: T
authors.lastname: Shiraki
authors.forename: Kyunghwa
authors.initials: K
authors.lastname: Park
authors.forename: Yuhichiro
authors.initials: Y
authors.lastname: Sakurai
authors.forename: Naohisa
authors.initials: N
authors.lastname: Tomosugi
authors.forename: Keiichi
authors.initials: K
authors.lastname: Kubota
mesh_terms: D000368:Aged; D023181:Antimicrobial Cationic Peptides / Q000096:biosynthesis*; D006528:Carcinoma, Hepatocellular / Q000378:metabolism*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D064451:Hepcidins; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D017403:In Situ Hybridization; D008113:Liver Neoplasms / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D010232:Paraffin / Q000737:chemistry; D016612:Paraffin Embedding / Q000379:methods; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods
chemical_list: D023181:Antimicrobial Cationic Peptides; C578702:HAMP protein, human; D064451:Hepcidins; D010232:Paraffin