| Project/Area Number |
16590962
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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 |
Hematology
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| Research Institution | Saitama Medical School |
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Principal Investigator |
YAGASAKI Fumiharu Saitama Medical School, Department of Hematology, Assistant Professor, 医学部, 講師 (40265418)
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| Co-Investigator(Kenkyū-buntansha) |
MATSUDA Akira Saitama Medical School, Department of Hematology, Associate Professor, 医学部, 助教授 (10219438)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2005: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2004: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | MDS / Histone H3 K4 / Histone H3 K9 / Methylation / ALU / LINE / Chromatin Immuno-precipation / monosomy 7 / monosomy7 / H3 K9 di-methylation / 複雑型染色体異常 / Alu |
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| Research Abstract |
Changes of DNA methylation and histone modifications have recently emerged as frequent events in hematopoietic malignancies. To clarify the global methylation status of histone H3 and its association with tumorigenesis of MDS, neutrophils separated from 10 consecutive MDS patients were analyzed by Western blotting using specific antibodies against di- or trimethylated H3-K9 and H3-K4. The methylation status was quantified as a Methylation Index (M.I.) using total H3 as an internal control and K562 cells as an external control. Compared with thresholds, which were defined as mean+SD of the M.I. in 9 healthy donors, global hyper di- and trimethylation of H3-K9 was found in 60.0% (M.I. range, 0.44-5.881;[threshold,1.573]), and 22.2% (0.227-1.639;[1.313]), and global hyper di- and trimethylation of H3-K4 in 44.4% (0.624-5.025;[1.656]) and 50.0% (0.756-26.114;[1.049]) of the patients, respectively. A significant correlation was found between global hyper-dimethylation of H3-K9 and -7 and complex chromosomal abnormalities including -7 (3/3:100%). Chromatin immunoprecipitation (ChIP) assays with dimethylated H3-K9 antibody followed by sequence analysis of immunoprecipitated DNA fragments of bone marrow mononuclear cells from a patient with -7 as the sole chromosomal abnormality revealed enrichment of repetitive elements including Alu (38.3%), LINEs (37.0%), MER (9.6%), LTR (4.1%) and MIR (4.1%) similar to previous reports. To identify methylated genes specific for MDS, ChIP subtraction assays are now in progress.
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