2003 Fiscal Year Final Research Report Summary
Elucidation of the Intracellular Signal Networks Involved in the Growth Arrest induced by Cell-cell Contact in Vascular Endothelial Cells
| Project/Area Number |
14570675
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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 |
Circulatory organs internal medicine
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| Research Institution | Kyushu University |
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Principal Investigator |
HIRANO Mayumi Kyushu University, Graduate School of Medical Sciences, Assistant Professor, 大学院・医学研究院, 助手 (80336031)
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| Co-Investigator(Kenkyū-buntansha) |
HIRANO Katsuya Kyushu University, Graduate School of Medical Sciences, Assistant Professor, 大学院・医学研究院, 講師 (80291516)
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| Project Period (FY) |
2002 – 2003
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| Keywords | vascular endothelial cells / cell-cell contact / p27^<Kip1> gene / promoter region / transcription activity / growth arrest |
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| Research Abstract |
1.The vascular endothelial cells ceased their growth upon formation of the tight cell-cell contact. The expression level of the cell cycle regulator p27^<Kip1> was upregulated during the contact-induced growth arrest. This up-regulation was found to be due to transcriptional up -regulation.
2.We have developed an assay system to determine the transcriptional regulatory element that responds to the formation of cell-cell contact in the cultured endothelia cells.
3.We obtained a porcine genomic clone containing a full-length p27^<Kip1> gene, and examined the promoter activity by using a 1500-nt up-stream region. The region -333to -247 nt was found to respond to the formation of homophilic cell-cell contact but not to the contact to HeLa cells. This promoter region is thus suggested to play an important role in up-regulating p27^<Kip1> expression during the contact-induced growth arrest.
4.We have developed a novel method to introduce protein into the cells with intact plasma membrane with a help of cell-penetrating peptide found in human immunodeficiency viral transcription factor Tat protein. By this method, proteins can be introduced in a quantitative and reversible manner. The time-specific transduction of the inhibitory peptide of RhoA revealed that the activity of-RhoA is required during the late G_1 phase of the cell cycle for the endothelial cells to progress to the S phase.
5.We found that co-transfection of the forkhead transcription factor AFX activated the p27^<Kip1> promoter.
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