Heat-Assisted Stretching of Paraffin Sections on Hot Plate Weakens Immunoreactivity of Orotate Phosphoribosyltransferase
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概要
- 論文の詳細を見る
Orotate phosphoribosyltransferase (OPRT) is the key enzyme for the phosphorylation of 5-fluorouracil (5-FU), the ratelimiting step for acquiring its anti-tumor effect. Since high enzyme activities and mRNA levels of OPRT are said to be associated with 5-FU chemosensitivity of cancer cells, the immunohistochemical demonstration of OPRT can be expected to contribute to the selection of patients who suffer from 5-FU-sensitive cancer. During a study for establishing the appropriate immunostaining condition using rabbit antiserum in formalin-fixed, paraffin-embedded sections of human cancer, we unexpectedly uncovered the fact that heat-assisted stretching of paraffin sections on a hot plate just after sectioning was critical for preserving OPRT immunoreactivity ; namely, stretching sections briefly at 70℃ or higher significantly reduced the immunoreactivity. Overnight drying of the sections in an oven at 37℃ or 60℃ did not influence the immunoreactivity, but pretreatments, including 0.2% trypsin, 0.002% proteinase K, and pressure cooking in 10 mM citrate buffer, pH 6.0 and pH 7.0, and 1 mM ethylenediaminetetraacetic acid solution, pH 8.0, seriously deteriorated the OPRT epitopes. The immunoreactivity was relatively resistant to overfixation, though weakened after fixation in formalin for four weeks. Xenografts with high OPRT enzyme activities showed distinct positive cytoplasmic staining, but those with low OPRT activities were equivocal. OPRT expression in routinely processed cancer tissues also provided valuable data.
- 日本組織細胞化学会の論文
著者
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Matsuoka Hiroshi
Department of Cardiology, Ehime Prefectural Imabari Hospital
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TSUTSUMI Yutaka
Department of Pathology, School of Medicine, Tokai University
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Tsutsumi Yutaka
Department Of Pathology Fujita Health University School Of Medicine
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Kamoshida Shingo
Department of Pathology, Fujita Health University School of Medicine
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Fukushima Masakazu
金沢大学がん研究所
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Fukushima Masakazu
Taiho Pharmaceutical Co.
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SAKURAI Yoichi
Department of Surgery, Fujita Health University School of Medicine
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Kamoshida Shingo
Department Of Pathology Fujita Health University School Of Medicine
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Sugimoto Yoshikazu
Tokushima Research Center Taiho Pharmaceutical
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Kamoshida S
Department Of Pathology Fujita Health University School Of Medicine
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Tsutsumi Yutaka
Department Of Internal Medicine Hakodate Municipal Hospital
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Sakamoto Nao
Department of Pathology, Fujita Health University School of Medicine
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Sakamoto Kazuki
Taiho Pharmaceutical Co.
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Sugimoto Yoshikazu
Taiho Pharmaceutical Co.
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Sakamoto Nao
Department Of Pathology Fujita Health University School Of Medicine
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Sakurai Yoichi
Department Of Surgery Fujita Health University School Of Medicine
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Matsuoka H
Toshiba Corp. Kawasaki‐shi Jpn
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Matsuoka Hiroshi
Department Of Surgery Fujita Health University School Of Medicine
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Shiogama Kazuya
Tokushima Research Center Taiho Pharmaceutical
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Sakurai Yukihiro
Department Of Gastroenterology Kanto Medical Center Nippon Telegraph And Telephone East Corporation
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Matsuoka Hiroshi
Department Of Applied Chemistry Faculty Of Science & Technology Keio University
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Sakamoto Kazuki
Tokushima Research Center Taiho Pharmaceutical
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Matsuoka Hiroshi
Department Of Applied Chemistry Faculty Of Engineering Osaka University
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Sakurai Yoichi
Department of Macromolecular Science and Engineering, Graduate School of Science and Technology, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto 606-8585, Japan
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MATSUOKA Hiroshi
Department of Surgery, Fujita Health University School of Medicine
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