| Project/Area Number |
10470270
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Thoracic surgery
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| Research Institution | AKITA UNIVERSITY |
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Principal Investigator |
OGAWA Junichi Sch. Of Med., Akita Univ., Professor, 医学部, 教授 (20112774)
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| Co-Investigator(Kenkyū-buntansha) |
KAWAI Hideki Sch. Of Med., Akita Univ., Research Associate, 医学部, 助手 (20291271)
MINAMIYA Yoshihiro Sch. Of Med., Akita Univ., Associate Professor, 医学部, 助教授 (30239321)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥6,300,000 (Direct Cost: ¥6,300,000)
Fiscal Year 1999: ¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 1998: ¥3,100,000 (Direct Cost: ¥3,100,000)
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| Keywords | Cancer metastasis / organ specificity / endothelial cell / extracellular matrix / lung cancer |
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| Research Abstract |
Many cancers display characteristic organ colonization patterns that do not fit simple, anatomical-mechanical trapping theories of tumor cell dissemination. Organ preferences of metastatic spread appear to be mediated partly by the selective attachment of tumor cells to organ-specific, microvascular endothelium. To study these tumor cell-endothelial cell interactions in an efficient and reproducible manner, we made organ-specific endothelial cell modulated by the organ specific extracellular matrix. A549 lung adenocaricinoma cells prefer to adhere to the endothelial cell cultured on the lung extracellular matrix. We also investigated the tumor cell chemotaxis. Chemotaxis is a key step in the process of tumor cell invasion and metastasis. We studied the role of nonmuscle myosin light chain kinase (nm-MLCK) in tumor cell chemotaxis toward hepatocyte growth factor (HGF) using A549 cells. Our analysis entailed characterizing expression of nm-MLCK using Western blot; examining chemotaxis toward HGF in Boyden chambers equipped with 8 μm pore polycarbonate membranes; examining myosin II filament formation by immunolabeling cells with anti-myosin II antiserum; and examining myosin light chain (MLC) phosphorylation by myosin II immunoprecipitation, followed by immunoblotting with anti-phosphoserine and anti-phosphothreonine antibodies. To assess the dependency MLCK activity, experiments were performed in the presence and absence of the specific MLCK antagonist, ML-7. A549 cells were found to express a 214 kDa nm-MLCK, which in the presence of HGF, phosphorylated MLC on both serine and threonine residues. HGF also elicited myosin II filament formation and chemotaxis which was maximal at 24 h in the presence of 30 ng/mlHGF. All of the aforementioned reactions were inhibited by ML-7. Thus, it appears that nm-MLCK activity regulates HGF-induced A549 cell chemotaxis.
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