| Project/Area Number |
11670174
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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 |
Human pathology
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| Research Institution | Shiga University of Medical Science |
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Principal Investigator |
SUGIHARA Hiroyuki Shiga University of Medical Science, Faculty of Medicine, Associate Professor, 医学部, 助教授 (30171169)
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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 |
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 2000: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1999: ¥800,000 (Direct Cost: ¥800,000)
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| Keywords | CGH / DOP-PCR / DNA ploidy / microdissection / 胃癌 / FISH |
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| Research Abstract |
The purpose of thus project was to apply CGH analysis to diffuse-type human gastric carcinomas to disclose early chromosomal changes in this type of tumors. There are 2 essential methodological points that have to be overcome for this purpose : method for an enrichment of cancer cells and methodology for an approach to early chromosomal change from CGE data of the surgically resected tumors. Because of the difficulty of enrichment of cancer cells that is enough for CGH analysis, however, we tentatively changed the material to esophageal squamous eell carcinoma of which the cancer cells arc easy to be isolated in order to concentrate on the second point. Then we took multiple small samples from surgical materials and successfully got the CGH profile from the tumor and reference DNAs that were extracted, DOP-PCR-amplified and PCR-labeled with red or green fluorochromes. Using the G/R ratio profiles of CGH analysis and DNA ploidy data, we determined the absolute copy numbers of chromosomal parts and compare the copy numbers amoong multiple samples taken from individual tumors. This comparison enabled us to reconstruct the process of clonal evolution and divergence from the stemline to sidelines and to specify when genome duplication occurred. The DNA copy number changes are thus classified into the ones earlier and later than genome duplication. This approach may be useful to extract important earlier chromosome changes from numerous aberrations detected by CGH alone in individual tumors. In this project, we confirmed that the method for an inference of absolute copy number of chromosomal part from DNA ploidy and the shift size of the G/R ratio that had been established by using cell lines was really applicable to surgically resected primary tumors.
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