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Analysis of liver diesease related gene expression using subtraction cloning

Research Project

Project/Area Number 12670467
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Gastroenterology
Research InstitutionTokyo Medical and Dental University

Principal Investigator

ENOMOTO Nobuyuki  Tokyo Medical and Dental University, Department of Gastroenterology and Hepatology, assistant professor, 医学部・附属病院, 講師 (20251530)

Co-Investigator(Kenkyū-buntansha) KUROSAKI Masayuki  Tokyo Medical and Dental University, Department of Gastroenterology and Hepatology, instructor, 医学部・附属病院, 助手 (10280976)
Project Period (FY) 2000 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2001: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 2000: ¥2,100,000 (Direct Cost: ¥2,100,000)
Keywordssubtraction / hepatoma / chronic hepatitis / autoimmune hepatitis / primary biliary cirrhosis
Research Abstract

The genetic basis of hepatocellular carcinoma (HCC) has not yet been fully understood. Although various methods have been developed to detect differentially expressed genes in malignant diseases, efficient analysis from clinical specimens is generally difficult to perform due to the requirement of a large amount of samples. In the present study, we analyzed differentially expressed genes with a small amount of human HCC samples using suppression subtractive hybridization (SSH). Total RNA were obtained from the hepatitis C virus-associated HCC and adjacent to non-HCC liver tissues. cDNA was synthesized using modified RT-PCR, and then tester cDNA was ligated with two different kinds of adaptors and hybridized with an excess amount of driver cDNA. Tester specific cDNA was obtained by suppression PCR and the final PCR product was subcloned and sequenced. Seven known genes (focal adhesion kinase, deleted in colon cancer, guanine binding inhibitory protein a, glutamine synthetase, ornithine aminotransferase, M130, and pepsinogen C) and two previously unknown genes as being overexpressed in HCC, and 1 gene (decorin) as suppressed in HCC. Quantitative analysis of gene expression using quantitative RT-PCR demonstrated the differential expression of these genes in the original and other HCC samples. These findings demonstrated that it is possible to identify the previously unknown, differential gene expression from a small amount of clinical samples. Information about such alterations in gene expression could be useful for elucidating the genetic events in HCC pathogenesis, developing the new diagnosic markers, or determining novel therapeutic targets. Using similar technique, we found overexpression of IP10 in autoimmune hepatitis and chronic hepatitis C. In PBC, several mitochondrial genes are up-regulated.

Report

(3 results)
  • 2001 Annual Research Report   Final Research Report Summary
  • 2000 Annual Research Report
  • Research Products

    (6 results)

All Other

All Publications (6 results)

  • [Publications] Miyasaka Y: "Analysis of differentially expressed genes in human hepatocellular carcinoma using suppression subtractive hybridization"Br J Cancer. 85. 228-234 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Nagayama K: "Overexpression of interferon gamma-inducible protein 10 in the liver of patients with type I autoimmune hepatitis identified by suppression subtractive hybridization"Am J Gastroenterol. 96. 2211-2217 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Miyasaka Y, Enomoto N, Nagayama K, Izumi N, Marumo F, Watanabe M, Sato C.: "Analysis of differentially expressed genes in human hepatocellular carcinoma using suppression subtractive hybridization"Br J Cancer. 85. 228-34 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Nagayama K, Enomoto N, Miyasaka Y Kurosaki M, Chen CH, Sakamoto N, Nakagawa M, Sato C, Tazawa J, Ikeda T, Izumi N, Watanabe M.: "Overexpression of interferon gamma-inducible protein 10 in the liver of patients with type I autoimmune hepatitis identified by suppression subtractive hybridization"Am J Gastroenterol. 96. 2211-7 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Miyasaka Y: "Analysis of differentially expressed genes in human hepatocellular carcinoma using suppression subtractive hybridization"Br J Cancer. 85. 228-234 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Nagayama K: "Overexpression of interferon gamma-inducible protein 10 in the liver of patients with type I autoimmune hepatitis identified by suppression subtractive hybridization"Am J Gastroenterol. 96. 2211-2217 (2001)

    • Related Report
      2001 Annual Research Report

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Published: 2000-04-01   Modified: 2016-04-21  

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