| Project/Area Number |
11671980
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Surgical dentistry
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| Research Institution | GUNMA UNIVERSITY (2000)
Tokyo Medical and Dental University (1999) |
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Principal Investigator |
NEGISHI Akihide GUNMA UNIVERSITY, DEPARTMENT OF ORAL AND MAXILLOFACIAL SURGERY, SCHOOL OF MEDICINE, ASSISTANT PROFESSOR, 医学部, 講師 (60270914)
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| Co-Investigator(Kenkyū-buntansha) |
吉田 光明 東京医科歯科大学, 難治疾患研究所, 助手 (60182789)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2000: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1999: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | ORAL CANCER / SQUAMOUS CELL CARCINOMA / GENETICS / CHROMOSOME / MICROCELL FUSION / TUMOR SUPPRESSOR GENE / 細胞 |
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| Research Abstract |
1)Construction of A9 cells containing subchromosomal transferable fragments(STFs)derived from the normal human chromosome 3.
A9(neo3)cells, containing normal human chromosome 3 tagged with pSV2neo, were irradiated with doses of 2-12 Gy of X-rays. Irradiated microcells were fused to A9 cells, and G418-resistant A9(STF)clones were selected in DMEM supplemented with 10% CS and 0.8mg/ml of G418. We have constructed 16 A9(STF)clones containing partial fragment derived from normal human chromosome 3. To identify the human sequence in the A9 clones containing STFs, we performed Q-banding analysis and chromosome painting with the human chromosome 3 specific probe. To localize the isolated STFs, DNA samples prepared from all 16 A9(STF)clones were assayed using 54 microsatellite markers on chromosome 3.
2)Tumorigenicity of the parental cell line and A9(STF)clones.
The tumorigenicity of the parental A9 cells and 6 different A9(STF)clones was examined in nude mice. The parental A9 cells formed progressively growing tumor in all the sites of nude mice within 3 to 5 weeks after subcutaneous inoculation. On the other hand, only one microcell hybrid ; A9(neo3B)cells failed to form tumors even more than 6 month after subcutaneous injection. Another five A9(STF)clones produced tumor in nude mice like the parental A9 cells. These data suggested that the critical tumor suppressor gene(s)which may contribute to the development of tumor in nude mice on human chromosome 3 was expressed only in the A9(neo3B) cells.
3)Identification of the critical tumor suppressor gene(s)on human chromosome 3 using suppression subtractive hybridization(SSH).
We are now trying to identify the critical tumor suppressor gene(s), which was expressed in the suppressed clone, not in the unsuppressed clones, on human chromosome 3 using SSH method.
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