| Project/Area Number |
63570929
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
外科・放射線系歯学
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| Research Institution | Tokyo Medical & Dental University |
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Principal Investigator |
SHIOIRI Shigeaki Tokyo Med. & Dent. University, 1st Dept. of Oral & Maxillofacial Surgery, Lecturer, 歯学部第一口腔外科, 講師 (90124693)
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| Co-Investigator(Kenkyū-buntansha) |
TSUCHIDA Nobuo Tokyo Med. & Dent. University, Dept. of Microbiology, Professor, 歯学部口腔細菌, 教授 (60089951)
SHIODA Shigetoshi Tokyo Med. & Dent. University, 1st Dept. of Oral & Maxillofacial Surgery, Profes, 歯学部第一口腔外科, 教授 (70041267)
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| Project Period (FY) |
1988 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1989: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1988: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Human Oral Cancer / p21 / Rb gene / transcriptional controlling element / c-H-ras-1 / c-myc / ヒトロ腔癌 / 癌遺伝子 / RB遺伝子 / 活性化ras / myc / 遺伝子導入 |
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| Research Abstract |
1) The ras p21 expression in human oral carcinoma tissue specimens was studied by immunohistochemistry with monoclonal antibodies against p2l. Not only malignant tumors, but also benigns were stained positively, and the staining intensities were relative. Therefore, it seemed to be impossible to evaluate the clinical malignancy. In addition, the expressions of Rb gene mRNA was investigated in seven human oral carcinoma cell lines and found to be normal in all of the cell lines.
2) The 2.9kb Sac I fragment of EJ c-H-ras-1 with 4 coding exons but without its own 5' promoter region and the VTR at 3' side was linked to tumor viruses's promoter/enhancer or only enhancer region at the 3' end in the opposite direction. These constructions in pUC18 plasmid were transfected onto NIH3T3 cells and found to induce foci with efficiencies of about 10% of those of the corresponding constructs in which the promoter/enhancer fragments were linked to the fragment in the same direction, while the transforming efficiency of the truncated c-H-ras-1 alone was less than 1%. The results suggests that viral enhancers increase transforming activity of the truncated c-H-ras-1. To further study the enhancement, upstream deletions were introduced into the truncated e-H-ras-1 linked to Moloney LTR in the opposite direction, and deletion plasmids were transfected onto NIH3T3 cells. Deletions from the upstream SacI site (-615) to around -150 from the initiation codon did not significantly lower the transforming activities, while the transforming activities of deletion mutants from -15O to -50 were low, suggesting that the controlling element may reside within this region. We examined also transforming activity of the truncated c-H-ras-1 by cotransfecting with c-myc linked to MoMSV LTR, in 3Y1 cells. The number of induced foci increased several fold, suggesting that c-myc enhances transforming activity of the truncated c-H-ras-1.
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