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Analysis of differentially expressed genes in human hepatocellular carcinoma using suppression subtractive hybridization

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...

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Autores principales: Miyasaka, Y, Enomoto, N, Nagayama, K, Izumi, N, Marumo, F, Watanabe, M, Sato, C
Formato: Texto
Lenguaje:English
Publicado: Nature Publishing Group 2001
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2364030/
https://www.ncbi.nlm.nih.gov/pubmed/11461082
http://dx.doi.org/10.1054/bjoc.2001.1901
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author Miyasaka, Y
Enomoto, N
Nagayama, K
Izumi, N
Marumo, F
Watanabe, M
Sato, C
author_facet Miyasaka, Y
Enomoto, N
Nagayama, K
Izumi, N
Marumo, F
Watanabe, M
Sato, C
author_sort Miyasaka, Y
collection PubMed
description 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 analysed 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 non-HCC liver tissues. cDNA was synthesized using modified RT-PCR, and then tester cDNA was ligated with 2 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. We identified 7 known genes (focal adhesion kinase, deleted in colon cancer, guanine binding inhibitory protein α, glutamine synthetase, ornithine aminotransferase, M130, and pepsinogen C) and 2 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. © 2001 Cancer Research Campaign http://www.bjcancer.com
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spelling pubmed-23640302009-09-10 Analysis of differentially expressed genes in human hepatocellular carcinoma using suppression subtractive hybridization Miyasaka, Y Enomoto, N Nagayama, K Izumi, N Marumo, F Watanabe, M Sato, C Br J Cancer Regular Article 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 analysed 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 non-HCC liver tissues. cDNA was synthesized using modified RT-PCR, and then tester cDNA was ligated with 2 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. We identified 7 known genes (focal adhesion kinase, deleted in colon cancer, guanine binding inhibitory protein α, glutamine synthetase, ornithine aminotransferase, M130, and pepsinogen C) and 2 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. © 2001 Cancer Research Campaign http://www.bjcancer.com Nature Publishing Group 2001-07 /pmc/articles/PMC2364030/ /pubmed/11461082 http://dx.doi.org/10.1054/bjoc.2001.1901 Text en Copyright © 2001 Cancer Research Campaign https://creativecommons.org/licenses/by/4.0/This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material.If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit https://creativecommons.org/licenses/by/4.0/.
spellingShingle Regular Article
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
title Analysis of differentially expressed genes in human hepatocellular carcinoma using suppression subtractive hybridization
title_full Analysis of differentially expressed genes in human hepatocellular carcinoma using suppression subtractive hybridization
title_fullStr Analysis of differentially expressed genes in human hepatocellular carcinoma using suppression subtractive hybridization
title_full_unstemmed Analysis of differentially expressed genes in human hepatocellular carcinoma using suppression subtractive hybridization
title_short Analysis of differentially expressed genes in human hepatocellular carcinoma using suppression subtractive hybridization
title_sort analysis of differentially expressed genes in human hepatocellular carcinoma using suppression subtractive hybridization
topic Regular Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2364030/
https://www.ncbi.nlm.nih.gov/pubmed/11461082
http://dx.doi.org/10.1054/bjoc.2001.1901
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