Gastroenterology

Gastroenterology

Volume 120, Issue 4, March 2001, Pages 955-966
Gastroenterology

Liver, Pancreas, and Biliary Tract
Differential gene expression between chronic hepatitis B and C hepatic lesion

https://doi.org/10.1053/gast.2001.22468Get rights and content

Abstract

Background & Aims: Complementary DNA (cDNA) microarray technology allows simultaneous expression analysis of hundreds to thousands of genes. We applied the cDNA microarray technique to clarify gene expression profiles in chronic viral hepatitis tissue lesions. Methods: We made cDNA microarrays consisting of 1080 human cDNAs and analyzed gene expression using labeled cDNAs prepared from 6 normal, 12 chronic hepatitis B, and 14 chronic hepatitis C liver tissues. Relative expression ratios of individual genes were obtained by comparing hybridization of Cy5-labeled cDNAs from chronic hepatitis lesions and Cy3-labeled cDNA from normal liver tissue. Results: Hierarchical clustering analysis of the gene expression profiles in 26 patients showed that the patients were clustered into 2 groups with respect to similarities in differentially expressed genes. Hepatitis B and C virus infection, but not age, sex, or histology of hepatitis, were significant factors determining clustering (P < 0.05). In hepatitis B tissue lesions, genes involved in inflammation were predominant, whereas in hepatitis C, expression of anti-inflammatory response genes was relatively dominant. Conclusions: These findings shed new light on the possible differential molecular mechanisms in the pathogenesis of hepatitis caused by hepatitis B virus and hepatitis C virus infection, from which hepatocellular carcinoma frequently develops.

GASTROENTEROLOGY 2001;120:955-966

Section snippets

Patients

The subjects were 12 patients with chronic hepatitis B, 14 patients with chronic hepatitis C, and 6 patients who showed no clinical signs of hepatitis in Kanazawa University Hospital, Japan, between 1997 and 1999 (Table 1).The last 6 patients were seronegative for either hepatitis B surface antigen (HBsAg) or HCV antibody and had liver function values within the normal limits. Tissue samples from these 6 patients were surgically obtained from the noncancerous parts of the liver complicated with

Establishment of a cDNA microarray technique for gene expression profiling in liver biopsy samples

Because the amount of mRNA recovered from small biopsy samples was not sufficient for cDNA microarray analysis, aRNA amplification of samples was necessary. To evaluate the sensitivity and reproducibility of cDNA microarray profiling using aRNA amplification, we performed preliminary experiments on a small scale, with 64 selected genes and mRNA from tissue culture cell lines. We could amplify RNA using 20 ng of mRNA as a template from THLE5-b and Huh-7 cells and used the amplified products for

Discussion

As a breakthrough technology for the development of modern functional research in human genome-based science, cDNA microarray has become one of the most powerful and sensitive techniques that can be broadly applied to both basic and clinical research. The cDNA microarray method makes it practical to quantitatively measure expression levels of hundreds to thousands of genes in parallel and has been used successfully to observe alterations and variations of gene expression in a variety of tissue

Acknowledgements

The authors thank Dr. Chris Barry for practical and productive technical advice on the microarray technique; Dr. Yasunari Nakamoto for discussion concerning the manuscript; Dr. Katsuhisa Inamura, Dr. Kazunori Kawaguchi, Dr. Katsuhiko Higuchi, Akemi Nakano, Junko Kitagawa, and Masami Ueda for excellent technical assistance; and Professor Seishi Murakami, Associate Professor Motoaki Ohtsubo, and Dr. Nobuyuki Itoh for providing cDNA clones.

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    Address requests for reprints to: Masao Honda, M.D., Ph.D., First Department of Internal Medicine, School of Medicine, Kanazawa University, Takara-Machi 13-1, Kanazawa, 920-8641, Japan. e-mail: [email protected]; fax: (81) 76-234-4250.

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