Investigation of the novel molecular target that determine the malignant character of pancreatic cancer cells.
| Project/Area Number |
15590621
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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 |
Gastroenterology
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| Research Institution | The University of Tokyo |
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Principal Investigator |
KOMATSU Yutaka The University of Tokyo, Faculty of Medicine, Assistant, 医学部附属病院, 助手 (90301100)
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| Co-Investigator(Kenkyū-buntansha) |
TATEISHI Keisuke The University of Tokyo, Faculty of Medicine, Assistant, 医学部附属病院, 助手
KAWAKAMI Takayuki The University of Tokyo, Faculty of Medicine, Medical Staff, 医学部附属病院, 医員 (60376457)
OTUKA Motoyuki The University of Tokyo, Faculty of Medicine, Medical Staff, 医学部附属病院, 医員
IJICHI Hieaki The University of Tokyo, Faculty of Medicine, Medical Staff, 医学部附属病院, 医員
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2004: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2003: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | smad4 / proteomics / microarray / Smad4 / 膵癌 / シグナル伝達 / トランスクリプトーム解析 / プロテオーム解析 |
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| Research Abstract |
The transforming growth factor-beta (TGF-beta)-Smad signaling pathway inhibits the growth of human epithelial cells and plays a role in tumor suppression. We succeeded in establishing Smad4 knockdown (S4KD) pancreatic cancer cell lines using the stable RNA interference (RNAi) method. The S4KD and control cells were stimulated with TGF-beta and analyzed using a cDNA microarray that contained 3756 genes, in order to screen for target molecules downstream of TGF-beta. The microarray analysis revealed that 187 S4KD genes and 155 genes in the control cells were regulated immediately upon TGF-beta stimulation. Quantitative RT-PCR analysis on several of these genes produced results that corroborated the outcome of the microarray analysis. Most of the genes in the S4KD and control cells identified by the array differed, which suggests signaling pathways that differ according to Smad4 status. Of the identified genes, 246 have not been reported previously as genes that lie downstream of TGF-beta. Genes that are involved in cell proliferation, adhesion, and motility were found to be regulated differentially with respect to S4KD and control cells. Cell migration induced by TGF-beta was inhibited in the S4KD cells, which might be associated with a different regulation of integrin beta7. In a while, we compared the proteomic changes with TGF-beta stimulation using two-dimensional gel electrophoresis (2-DE) and mass spectrometry. We identified five proteins that were up-regulated and seven proteins that were down-regulated ; 10 of them were novel targets for TGF-beta. These proteins function in processes such as cytoskeletal regulation, cell cycle, and oxidative stress. Introducing siRNA-mediated gene silencing into proteomics revealed a novel TGF-beta signal pathway that did not involve Smad4.
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Report
(3 results)
Research Products
(14 results)
