| Project/Area Number |
18591441
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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 |
General surgery
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| Research Institution | Kumamoto University |
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Principal Investigator |
KAMOHARA Hidenobu Kumamoto University, University Hospital, Assistant (90398222)
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| Co-Investigator(Kenkyū-buntansha) |
BABA Hideo Kumamoto University, Faculty of Medical and Pharmaceutical Sciences, Professor (20240905)
IWASE Hirotaka Kumamoto University, Faculty of Medical and Pharmaceutical Sciences, Professor (40211065)
IYAMA Kenichi Kumamoto University, University Hospital, Associate Professor (10040536)
ISHIKO Takatoshi Kumamoto University, University Hospital, Medical Staff (00343351)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,920,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥420,000)
Fiscal Year 2007: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2006: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | Receptor tyrosine kinase / Collagen / Cancer cells / Stromal i.nvasion(M.etastasis) / RNA interference / RNA平渉 |
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| Research Abstract |
Discoidin domain receptor 1 (DDR1) is a tyrosine kinase receptor, and its ligand is collagens. To identify DDR1 biological function for cancer cells, we made DDR1 stably knock down cells (K/D) using retro viral system in MCF7 cells (mammary carcinoma cell lines) expresses DDR1 highly. K/D lost cell-to-cell contact and extended pesudopods in cellular morphology. This cellular change was observed significantly in the leading edge of cell cluster with collagen. K/D promoted the biological activity of cells growth, migration, and invasion in comparison with mock (retro viral vector only expressing cells). PI3K inhibitor, LY294002 and wortmmanin, suppressed cells growth significantly in K/D. Additionally, neutralizing 131-integrin antibodyin suppressed cells growth in K/D. Phosphorylation of Akt (Ser473 and Thr308) was enhanced in the prence of collaen in K/D. FAK and CAS were phosphorylated strongly in K/D, but DDR1 itself was not in both K/D and mock. Additionally, western blotting and immunoprecipitation with anti-DDR1, FAK or CAS IgG indicated cooperation among them under the stimulation of collagen. Phophrylation of FAK and CAS was enhanced by collagen in K/D. DDR1 has two major isoform, DDR1 a and DDR1b. DDR1 a deleted 37 amino acids in intracellular domain by alternative splicing. To reveal the difference of these isoforms, we made DRla or DDR1b overexpressing MDA-MB-23l cells which expressed DDR1 a mainly. MCF7 expressed DDRb isoform. Observations of DDRlb overexpressing MDA-MB-231 cells suggested that DDR1b suppressed cells invasion significantly. In summary, our suppression of DDR1 study suggested that DDR1 negatively regulated cancer progression with collagen. Futurely, DDR1 isoforms could be indicators of degree of malignancy and molecular targets of cancer therapy.
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