Brief Article
Copyright ©2012 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Gastroenterol. Jul 28, 2012; 18(28): 3727-3731
Published online Jul 28, 2012. doi: 10.3748/wjg.v18.i28.3727
Analysis of hepcidin expression: In situ hybridization and quantitative polymerase chain reaction from paraffin sections
Yuhki Sakuraoka, Tokihiko Sawada, Takayuki Shiraki, Kyunghwa Park, Yuhichiro Sakurai, Naohisa Tomosugi, Keiichi Kubota
Yuhki Sakuraoka, Tokihiko Sawada, Takayuki Shiraki, Kyunghwa Park, Yuhichiro Sakurai, Keiichi Kubota, Second Department of Surgery, School of Medicine, Dokkyo Medical University, Kitakobayashi 880, Mibu, Shimotsuga, Tochigi 321-0293, Japan
Naohisa Tomosugi, Proteomics Research Unit, Division of Advanced Medicine, Medical Research Institute, Kanazawa Medical College, Daigaku 1-1, Uchinada, Kanazawa 920-0293, Japan
Author contributions: Sakuraoka Y and Sawada T performed the study and wrote the paper; Shiraki T, Park K, and Sakurai Y assisted with the study; Sawada T designed the study; Tomosugi N and Kubota K assisted in the study and reviewed the paper.
Supported by A research grant from the Biomarker Society
Correspondence to: Tokihiko Sawada, MD, PhD, Second Department of Surgery, School of Medicine, Dokkyo Medical University, Kitakobayashi 880, Mibu, Shimotsuga, Tochigi 321-0293, Japan. tsawada@dokkyomed.ac.jp
Telephone: +81-282-861111 Fax: +81-282-866317
Received: September 27, 2010
Revised: November 21, 2010
Accepted: May 12, 2012
Published online: July 28, 2012
Abstract

AIM: 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.

CONCLUSION: We have established novel methods for quantitative PCR for hepcidin using RNAs isolated from paraffin-embedded sections and in situ hybridization of HCC.

Keywords: Hepcidin; Expression; In situ hybridization; Immunohistochemistry; Real-time polymerase chain reaction