| Project/Area Number |
17590073
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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 |
Biological pharmacy
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| Research Institution | Tokyo University of Science |
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Principal Investigator |
SHIMIZU Takahisa Tokyo University of Science, Faculty of Pharmaceutical Sciences, Assistant, 薬学部, 助手 (30287479)
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| Co-Investigator(Kenkyū-buntansha) |
TAKEDA Ken Tokyo University of Science, Faculty of Pharmaceutical Sciences, Professor, 薬学部, 教授 (80054013)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2006: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2005: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Keywords | transcriptional regulation / induced differentiation / retinoic acid / leukemia / GM-CSF |
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| Research Abstract |
We reported previously that treatment with all-trans retinoic acid (ATRA) and granulocyte macrophage colony-stimulating factor (GM-CSF) induces synergistically differentiation of human myeloblastic leukemia ML-1 cells to granulocytes. To investigate the mechanism of transcriptional regulation of retinoic acid-responsive genes during the differentiation, we examined expression of the responsive genes, retinoic acid receptors (RARs), retinoid X receptors (RXRs) and the cofactors in ML-1 cells treated with ATRA and/or GM-CSF using Northern or Western blot analysis. We first showed that expression of C/EBP ε mRNA was induced synergistically by treatment with ATRA and GM-CSF. We next detected up-regulation of expression of RAR a mRNA and protein by GM-CSF and synergistic increase of expression of steroid receptor coactivator-3 (SRC3/AIB 1), which is known as activator of nuclear receptors including RARs, mRNA and protein by both reagents. These changes were also detected in other myeloid leukemia cell lines (THP-1 and KG-1) that showed a synergistic effect similar to that seen in ML-1 cells in response to ATRA and GM-CSF. Furthermore, we revealed alteration of expression of genes regulating histone arginine methylation (PRMTs and PAD4). To investigate whether these alterations are associated with the differentiation, we examined effect of SRC3 siRNA on the differentiation in ML-1 cells treated with ATRA and GM-CSF. The inhibition of induced SRC3/AIB1 gene expression by SRC3 siRNA suppressed NBT reducing activity induced by treatment with both reagents. These results suggest that the synergistic induction of differentiation by ATRA and GM-CSF is associated with the increase of sensitiveness of cells to retinoic acid via up-regulation of expression of SRC3/AIB1 gene.
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