Related Experiment Video
Updated: May 20, 2026
Visualization, Quantification, and Mapping of Immune Cell Populations in the Tumor Microenvironment
Published on: March 25, 2020
Analysis of hepcidin expression: in situ hybridization and quantitative polymerase chain reaction from paraffin
Yuhki Sakuraoka1, Tokihiko Sawada, Takayuki Shiraki
1Second Department of Surgery, School of Medicine, Dokkyo Medical University, Kitakobayashi 880, Mibu, Shimotsuga, Tochigi 321-0293, Japan.
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.
