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Analysis of hepcidin expression: in situ hybridization and quantitative polymerase chain reaction from paraffin sections.

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

SUBMITTER: Sakuraoka Y 

PROVIDER: S-EPMC3406426 | biostudies-literature | 2012 Jul

REPOSITORIES: biostudies-literature

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Analysis of hepcidin expression: in situ hybridization and quantitative polymerase chain reaction from paraffin sections.

Sakuraoka Yuhki Y   Sawada Tokihiko T   Shiraki Takayuki T   Park Kyunghwa K   Sakurai Yuhichiro Y   Tomosugi Naohisa N   Kubota Keiichi K  

World journal of gastroenterology 20120701 28

<h4>Aim</h4>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).<h4>Methods</h4>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 we  ...[more]

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