Identification of Altered Protein Expression and Post-Translational Modifications in Primary Colorectal Cancer by Using Agarose Two-Dimensional Gel Electrophoresis

  • Takeshi Tomonaga
    1Molecular Diagnosis (F8) and Departments of
  • Kazuyuki Matsushita
    2Academic Surgery (M9), Graduate School of Medicine, Chiba University, Chiba, Japan, and
  • Seiko Yamaguchi
    1Molecular Diagnosis (F8) and Departments of
  • Masamichi Oh-Ishi
    3Laboratory of Biomolecular Dynamics, Department of Physics, Kitasato University School of Science, Kanagawa, Japan
  • Yoshio Kodera
    3Laboratory of Biomolecular Dynamics, Department of Physics, Kitasato University School of Science, Kanagawa, Japan
  • Tadakazu Maeda
    3Laboratory of Biomolecular Dynamics, Department of Physics, Kitasato University School of Science, Kanagawa, Japan
  • Hideaki Shimada
    2Academic Surgery (M9), Graduate School of Medicine, Chiba University, Chiba, Japan, and
  • Takenori Ochiai
    2Academic Surgery (M9), Graduate School of Medicine, Chiba University, Chiba, Japan, and
  • Fumio Nomura
    1Molecular Diagnosis (F8) and Departments of

抄録

<jats:title>Abstract</jats:title> <jats:p>Purpose: Although numerous proteome studies have been performed recently to identify cancer-related changes in protein expression, only a limited display of relatively abundant proteins has been identified. The aim of this study is to identify novel proteins as potential tumor markers in primary colorectal cancer tissues using a high-resolution two-dimensional gel electrophoresis (2-DE).</jats:p> <jats:p>Experimental Design: 2-DE using an agarose gel for isoelectric focusing was used to compare protein profiling of 10 colorectal cancer tissues and adjacent normal mucosa. Altered expression and post-translational modification of several proteins were examined using Western blot analysis and immunohistochemistry.</jats:p> <jats:p>Results: Ninety-seven proteins of 107 spots (90.7%) that were differentially expressed between matched normal and tumor tissues were identified by mass spectrometry. Among them, 42 unique proteins (49 spots) significantly increased or decreased in the tumors. They include eukaryotic translation initiation factor 4H, inorganic pyrophosphatase, anterior gradient 2 homologue, aldolase A, and chloride intracellular channel 1, whose elevated expression in tumor tissues was confirmed by Western blot analysis and immunohistochemistry. Interestingly, only isoform 1 of two transcript variants of eukaryotic translation initiation factor 4H was greatly up-regulated in most of the tumor tissues. Moreover, post-translational modifications of the prolyl-4-hydroxylase β subunit and annexin A2 also were identified.</jats:p> <jats:p>Conclusions: We identified several novel proteins with altered expression in primary colorectal cancer using agarose 2-DE. This method is a powerful technique with which to search for not only quantitative but also qualitative changes in a biological process of interest and may contribute to the deeper understanding of underlying mechanisms of human cancer.</jats:p>

収録刊行物

  • Clinical Cancer Research

    Clinical Cancer Research 10 (6), 2007-2014, 2004-03-15

    American Association for Cancer Research (AACR)

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