Sakuraoka Y, Sawada T, Shiraki T, Park K, Sakurai Y, Tomosugi N, Kubota K. Analysis of hepcidin expression: In situ hybridization and quantitative polymerase chain reaction from paraffin sections. World J Gastroenterol 2012; 18(28): 3727-3731 [PMID: 22851866 DOI: 10.3748/wjg.v18.i28.3727]
Corresponding Author of This Article
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
Article-Type of This Article
Brief Article
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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.
