| Project/Area Number |
01440030
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| Research Category |
Grant-in-Aid for General Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
Human pathology
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| Research Institution | Fukui Medical School |
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Principal Investigator |
FUKUDA Masaru Fukui Medical School, Dept. Path. Prof., 医学部, 教授 (60079720)
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| Co-Investigator(Kenkyū-buntansha) |
NORIKI Sakon Fukui Med. School, Dept. Path. Ass., 医学部, 助手 (30228374)
MIYOSHI Norio Fukui Med. School, Dept. Path. Ass., 医学部, 助手 (40209961)
SUGIHARA Hiroyuki Fukui Med. School, Dept Path. Ass., 医学部, 助手 (30171169)
NAKANISHI Kazuo Fukui Med. School, Dept Path. Ass. Prof., 医学部, 助教授 (10094434)
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| Project Period (FY) |
1989 – 1991
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| Project Status |
Completed (Fiscal Year 1991)
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| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1991: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1990: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Cancer markers / DNA instability / AO-fluorescent staining / Anti-single-stranded DNA antibody / STA-glycoprotein cDNA / Anti-STA-glycoprotein antibody / Active oxygens / Cancer diagnosis / AO蛍光染色 / STA結合糖蛋白質cDNA / STA結合糖蛋白質 / in situ hybridization / Photon counting / 悪性腫瘍マ-カ- / DNA損傷 / アクリジンオレンジ / 糖蛋白 / モノクロ-ナル抗体 / ケミルミ |
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| Research Abstract |
The aim of the present research work is to develop new cancer markers for the daily diagnosis of malignancy based on so far discovered new-characters of cancerous cells, as follow ; (1) most malignant tumor cells have more unstable nuclear DNA than comparable benign tumor cells and normal-cells, (2) malignant tumor cells bear a common cytoplasmic glycoprotein stainable with STA by lectin histochemistry, and (3) malignant cells leberate massive activeoxygens. Acid hydrolysis with 2N HCI at 30ﾟC forls min after RNase treatment was found to induce selective DNA denaturation of cancer cells in ordinary paraffin-embedded sections, and the single-stranded DN, A was differentially stained immunohistochemically with anti-singlestranded polyclonal antibody. The cDNA. libray of human thyroid cancer was incorporated to E. coli, and the CDNA for STA-positive-glycoprotein and STA-positive mutant B. coli clones-were obtained, and the poly-clonal antibody against for the glycoprotein was also obtained. By using the CDNA and antibody, differential staining of malignant tumor cells was successfully attained by in-situ-hybridization or immunohistochemistry, respectively. The liberation of active oxygen was detected from a Sransplanted squamous cell carcinoma cells of mouse by a photon-counting imaging system (VIM, Hamamatsu Photnics) using a chemiluminescence sensitizer MCLA. Especially the former two methods can be readily used for daily pathological diagnosis of malignancy as reliable cancdr markers.
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