2002 Fiscal Year Annual Research Report

線溶系因子による細胞運動調節機構の細胞表面分子間反応の可視化による解析

Research Project

Project/Area Number	13670040
Research Institution	Hamamatsu University School of Medicine
Principal Investigator	浦野哲盟浜松医科大学, 医学部, 教授 (50193967)
Co-Investigator(Kenkyū-buntansha)	最上秀夫浜松医科大学, 医学部, 助教授 (90311604) 山本清二浜松医科大学, 光量子医学研究センター, 助教授 (60144094) 井原勇人浜松医科大学, 医学部, 助手 (00223298) 鈴木優子浜松医科大学, 医学部, 助手 (20345812)
Keywords	細胞接着 / 細胞移動 / ウロキナーゼ受容体 / ウロキナーゼ / PAI-1 / PAI-2 / GFP
Research Abstract	urokinase type plasminogen activator(uPA)は癌細胞表面や新生血管内皮細胞上の特異受容体(uPAR)に結合しており、そのインヒビターであるPA inhibitor 1(PAI-1)も腫瘍組織に多く存在する。本研究はこれらの機能蛋白の分子間相互反応を可視化し、腫瘍細胞の移動や血管新生時の細胞運動における役割を明らかにすることを目的とする。本年度の研究において以下の結果が得られた。 uPAR及びuPAを発現しているhuman sarcoma cell lineであるHT1080はfibronectin(FN)に比較して、Vitronectin(Vn)-及びCollagen IV(Col)-coatのdishによく接着した。PAI-1はVnと結合することにより、HT1080のVnへ接着を濃度依存性に阻害したが、FNあるいはColへの接着は阻害しなかった。また接着能を変えることにより、PAI-1はVnからColへの細胞移動を促進した。uPAはPAI-1と結合し、PAI-1のVn結合能を喪失させた。これによりuPAは上記のPAI-1による接着能の変化及び遊走能の変化を喪失させた。特異抗体を用いて、HT1080のαIIβVintegrinとVnの接着をPAI-1及びuPAが修飾する事実が確認できた。Green Fluorescent Protein(GFP)標識uPARの作成とHT1080への導入に成功した。現在FITC標識uPAも併用してエヴァネッセント蛍光顕微鏡で細胞移動時のこれらの標識蛋白の局在の変化を解析中である。

Research Products

(10 results)

All Other

All Publications (10 results)

[Publications] Pawlak R,. et al.: "The differential effect of angiotensin II and angiotensin 1-7 on norepinephrine, epinephrine, and dopamine concentrations in rat hypothalamus : the involvement of angiotensin receptors"Brain Res Bull.. 54・6. 689-694 (2001)
[Publications] Yoshino A. et al.: "Enhanced secretion of tissue plasminogen activator by simultaneous use of retinoic acid and ascorbic acid from tissue cultured gastroepiploic artery"Life Sciences. 70. 1461-1470 (2002)
[Publications] Aoki K.et al.: "Elevation of plasma free PAI-1 levels as an integrated endothelial response to severe burs"Burns. 27. 569-575 (2001)
[Publications] Pawlak R. et al.: "Rapid, specific and active site-catalyzed effect of tissue-plasminogen activator on hippocampus-dependent learning in mice"Neuroscience. 113/4. 995-1001 (2002)
[Publications] Yoshida M. et al.: "L-158,809 and (D-Ala7)-angiotensin I/II (1-7) decrease PAI-1 release from human umbilical vein endothelial cells"Thromb. Res.. 105/6. 531-536 (2002)
[Publications] Nagai N. et al.: "Tissue-type plasminogen activator has paradoxical roles in focal cerebral ischemic injury by thrombotic middle cerebral artery occlusion with mild or severe photochemical damage in mice"J Cereb Blood Flow Metab.. 22. 648-651 (2002)
[Publications] Suzuki Y, et al.: "Dietary supplementation with fermented soybeans suppresses intimal thickening"Nutrition. (in press).
[Publications] Sakata K. et al.: "Effects of a longterm pharmacological interruption of the rennin-angiotensin system on the fibrinolytic system in essential hypertension"Pathophysiology of Haemostasis and Thrombosis. 32. 67-75 (2002)
[Publications] Takahashi H. et al.: "5-HT4 receptors in the hippocampus modulate rat locomotor activity"Hippocampus. 12. 304-310 (2002)
[Publications] Mogami H. et al.: "Decoding of short-lived Ca2+influx signals into long-term substrate phosphorylation through activation of two distinct classes of protein kinase C"J. Biol. Chem. 278/11. 9896-9904 (2003)

2002 Fiscal Year Annual Research Report

線溶系因子による細胞運動調節機構の細胞表面分子間反応の可視化による解析

Principal Investigator

浦野 哲盟 浜松医科大学, 医学部, 教授 (50193967)

Research Products

[Publications] Pawlak R,. et al.: "The differential effect of angiotensin II and angiotensin 1-7 on norepinephrine, epinephrine, and dopamine concentrations in rat hypothalamus : the involvement of angiotensin receptors"Brain Res Bull.. 54・6. 689-694 (2001)

[Publications] Yoshino A. et al.: "Enhanced secretion of tissue plasminogen activator by simultaneous use of retinoic acid and ascorbic acid from tissue cultured gastroepiploic artery"Life Sciences. 70. 1461-1470 (2002)

[Publications] Aoki K.et al.: "Elevation of plasma free PAI-1 levels as an integrated endothelial response to severe burs"Burns. 27. 569-575 (2001)

[Publications] Pawlak R. et al.: "Rapid, specific and active site-catalyzed effect of tissue-plasminogen activator on hippocampus-dependent learning in mice"Neuroscience. 113/4. 995-1001 (2002)

[Publications] Yoshida M. et al.: "L-158,809 and (D-Ala7)-angiotensin I/II (1-7) decrease PAI-1 release from human umbilical vein endothelial cells"Thromb. Res.. 105/6. 531-536 (2002)

[Publications] Nagai N. et al.: "Tissue-type plasminogen activator has paradoxical roles in focal cerebral ischemic injury by thrombotic middle cerebral artery occlusion with mild or severe photochemical damage in mice"J Cereb Blood Flow Metab.. 22. 648-651 (2002)

[Publications] Suzuki Y, et al.: "Dietary supplementation with fermented soybeans suppresses intimal thickening"Nutrition. (in press).

[Publications] Sakata K. et al.: "Effects of a longterm pharmacological interruption of the rennin-angiotensin system on the fibrinolytic system in essential hypertension"Pathophysiology of Haemostasis and Thrombosis. 32. 67-75 (2002)

[Publications] Takahashi H. et al.: "5-HT4 receptors in the hippocampus modulate rat locomotor activity"Hippocampus. 12. 304-310 (2002)

[Publications] Mogami H. et al.: "Decoding of short-lived Ca2+influx signals into long-term substrate phosphorylation through activation of two distinct classes of protein kinase C"J. Biol. Chem. 278/11. 9896-9904 (2003)

浦野哲盟浜松医科大学, 医学部, 教授 (50193967)