2002 Fiscal Year Annual Research Report

モルフォーゲン活性を有する新規生理活性脂質S1Pの生理機能の分子生物学的解析

Research Project

Project/Area Number	13470008
Research Institution	Kanazawa University
Principal Investigator	多久和陽金沢大学, 医学系研究科, 教授 (60171592)
Co-Investigator(Kenkyū-buntansha)	吉岡和晃金沢大学, 医学系研究科, 助手 (80333368) 多久和典子金沢大学, 医学系研究科, 助手 (70150290) 杉本直俊金沢大学, 医学系研究科, 講師 (80272954)
Keywords	スフィンゴシン-1-リン酸 / EDG / AGR-16 / 受容体 / Rac / Rho / 細胞遊走
Research Abstract	Edg5ノックアウト(KO)マウス、S1P合成酵素Sphキナーゼトランスジェニック(Tg)マウス、S1P分解酵素S1PリアーゼTgマウスを作出した。また、これと平行してS1P・Edg受容体システムによる二方向性細胞運動制御の分子機構の解析を進めた。 Edg5 KOマウスはメンデル則にしたがって出生した後、離乳までに半数近くが死亡するほか、成長後も死亡率が高い。Edg5 KOマウスに10mmHg程度の血圧低下、Edg5 KO雌マウスに子宮形成不全、少仔傾向を認める他は明らかな形態形成の異常は見られない。現在死因を解析中である。Sphキナーゼ、S1PリアーゼTgマウスでは、創傷治癒ならびに血管傷害後内皮肥厚のそれぞれ促進、遅延を認めている。Edg5による化学遊走及びRacの抑制は、三量体G蛋白質G_<12/13>と、そのエフェクターRhoを介していることを初めて明らかにした。Edg3はEdg5と同様にG_<12/13>に共役してRhoを活性化するが、Edg5とは異なりRacを活性化して遊走を促進した。Edg3のこの作用は百日咳毒素前処理により消失し、逆にEdg5同様Rac活性ならびに遊走の抑制に転じたことから、Edg3では優勢なGi共役がG_<12/13>を介した抑制作用をマスクしている結果、Rac活性及び遊走が促進されることが判明した。またさらに、マウスB16メラノーマ細胞を用いたin vivo浸潤、尾静脈注射後肺転移のモデルシステムを活用し、癌の浸潤・転移を典型例とする生体内細胞運動が、発現するEdg受容体アイソフォーム依存性に特異的に制御されていることを初めて明らかにした。

Research Products

(16 results)

All Other

All Publications (16 results)

[Publications] N.Takuwa et al.: "Regulation of cell cycle molecules by the Ras effector system"Mol.Cell.Endocrinol.. 177(1-2). 25-33 (2001)
[Publications] Y.Takuwa et al.: "Subtype-Specific, Differential Activities of the EDG Family Receptors for Sphingosine-1-Phosphate, a Novel Lysophospholipid Mediateor"Mol.Cell.Endocrinol.. 177. 3-11 (2001)
[Publications] H.Mitui et al.: "The MEK1-ERK map kinase pathway and the PI 3-kinase-AKT pathway independently mediate anti-apoptotic signals in HEPG2 liver cancer cells"Int.J.Cancer.. 92. 55-62 (2001)
[Publications] S.Sakurada et al.: "Rho activation in excitatory agonist-stimulated vascular smooth muscle"Am.J.Physiol.(Cell Physiol). 281. C571-C578 (2001)
[Publications] Y.Banno et al.: "Involvement of phospholipase D in sphingosine 1-phosphate-induced activation of phosphatidylinositol 3-kinase and Akt in chinese hamster ovary cells overexpressing EDG3^*"J.Biol.Chem.. 276. 35622-35628 (2001)
[Publications] Y.Ryu et al.: "Sphingosine-1-Phosphate, a Platelet-Derived Lysophospholipid Mediator, Negatively Regulates Cellular Rac Activity and Cell Migration in Vascular Smooth Muscle Cells"Circ Res.. 90. 325-332 (2002)
[Publications] Y.Takuwa et al.: "The Edg G protein-coupled receptors for lysophospholipids : Their signaling prope rties and biological activities"J.Biochem.. 131. 767-771 (2002)
[Publications] Y.Takuwa et al.: "Subtype-specific differential regulation of Rho family G proteins and cell migration by the Edg family sohingosine-1-phosphate receptors"Biophys.Biochim.Acta.. 1582(1-3). 112-120 (2002)
[Publications] T.Ishibashi et al.: "Inhibition of Rho/Rho-kinase signaling downregulates plasminogen activator inhibitor-1 synthesis in cultured human monocytes"Biophys.Biochim.Acta.. 1590(1-3). 123-130 (2002)
[Publications] K.Nagata et al.: "Rho/Rho-kinase is involved in the synthesis of tissue factor in human monocytes"Atherosclerosis.. 163(1). 39-47 (2002)
[Publications] Y.Banno et al.: "Involvement of phospholipase D in insulin-like growth factor-I-induced activation of extracellular signal-regulated kinase, but not phosphatidylinositol 3-kinase or Akt, in chinese hamster ovary cells"Biochem.J.. 369. 363-368 (2003)
[Publications] H.Ikeda et al.: "Antiproliferative action of sphingosine 1-phosphate in rat hepatocytes involves activation of Rho via Edg-5"Gastroenterology.. (In press). (2003)
[Publications] K.Ito et al.: "Mechanisms recponsible for Ca^<2+> sensitization of prostaglandin F2α-induced contraction of vascular smooth muscle"J.Physiol.. (In press). (2003)
[Publications] N.Sugimoto et al.: "Inhibitory and stimulatory regulation of Rac and cell motility by the G_<12/13>-Rho-and the G_1-pathwaysintegrated downstream of a single G protein coupled sphingosine-1-phosphate receptor isoform"Mol.Cell.Biol.. 23. 1534-1545 (2003)
[Publications] D.Shida et al.: "Lysophosphatidic acid enhances metastatic potential of human colon carcinoma DLD1 cells through LPA1"Cancer Res.. (In press). (2003)
[Publications] 多久和陽: "血小板と生理活性脂質"尾崎由基男等(金芳堂). 182 (2002)

2002 Fiscal Year Annual Research Report

モルフォーゲン活性を有する新規生理活性脂質S1Pの生理機能の分子生物学的解析

Principal Investigator

多久和 陽 金沢大学, 医学系研究科, 教授 (60171592)

Research Products

[Publications] N.Takuwa et al.: "Regulation of cell cycle molecules by the Ras effector system"Mol.Cell.Endocrinol.. 177(1-2). 25-33 (2001)

[Publications] Y.Takuwa et al.: "Subtype-Specific, Differential Activities of the EDG Family Receptors for Sphingosine-1-Phosphate, a Novel Lysophospholipid Mediateor"Mol.Cell.Endocrinol.. 177. 3-11 (2001)

[Publications] H.Mitui et al.: "The MEK1-ERK map kinase pathway and the PI 3-kinase-AKT pathway independently mediate anti-apoptotic signals in HEPG2 liver cancer cells"Int.J.Cancer.. 92. 55-62 (2001)

[Publications] S.Sakurada et al.: "Rho activation in excitatory agonist-stimulated vascular smooth muscle"Am.J.Physiol.(Cell Physiol). 281. C571-C578 (2001)

[Publications] Y.Banno et al.: "Involvement of phospholipase D in sphingosine 1-phosphate-induced activation of phosphatidylinositol 3-kinase and Akt in chinese hamster ovary cells overexpressing EDG3^*"J.Biol.Chem.. 276. 35622-35628 (2001)

[Publications] Y.Ryu et al.: "Sphingosine-1-Phosphate, a Platelet-Derived Lysophospholipid Mediator, Negatively Regulates Cellular Rac Activity and Cell Migration in Vascular Smooth Muscle Cells"Circ Res.. 90. 325-332 (2002)

[Publications] Y.Takuwa et al.: "The Edg G protein-coupled receptors for lysophospholipids : Their signaling prope rties and biological activities"J.Biochem.. 131. 767-771 (2002)

[Publications] Y.Takuwa et al.: "Subtype-specific differential regulation of Rho family G proteins and cell migration by the Edg family sohingosine-1-phosphate receptors"Biophys.Biochim.Acta.. 1582(1-3). 112-120 (2002)

[Publications] T.Ishibashi et al.: "Inhibition of Rho/Rho-kinase signaling downregulates plasminogen activator inhibitor-1 synthesis in cultured human monocytes"Biophys.Biochim.Acta.. 1590(1-3). 123-130 (2002)

[Publications] K.Nagata et al.: "Rho/Rho-kinase is involved in the synthesis of tissue factor in human monocytes"Atherosclerosis.. 163(1). 39-47 (2002)

[Publications] Y.Banno et al.: "Involvement of phospholipase D in insulin-like growth factor-I-induced activation of extracellular signal-regulated kinase, but not phosphatidylinositol 3-kinase or Akt, in chinese hamster ovary cells"Biochem.J.. 369. 363-368 (2003)

[Publications] H.Ikeda et al.: "Antiproliferative action of sphingosine 1-phosphate in rat hepatocytes involves activation of Rho via Edg-5"Gastroenterology.. (In press). (2003)

[Publications] K.Ito et al.: "Mechanisms recponsible for Ca^<2+> sensitization of prostaglandin F2α-induced contraction of vascular smooth muscle"J.Physiol.. (In press). (2003)

[Publications] N.Sugimoto et al.: "Inhibitory and stimulatory regulation of Rac and cell motility by the G_<12/13>-Rho-and the G_1-pathwaysintegrated downstream of a single G protein coupled sphingosine-1-phosphate receptor isoform"Mol.Cell.Biol.. 23. 1534-1545 (2003)

[Publications] D.Shida et al.: "Lysophosphatidic acid enhances metastatic potential of human colon carcinoma DLD1 cells through LPA1"Cancer Res.. (In press). (2003)

[Publications] 多久和 陽: "血小板と生理活性脂質"尾崎由基男等(金芳堂). 182 (2002)

多久和陽金沢大学, 医学系研究科, 教授 (60171592)

[Publications] 多久和陽: "血小板と生理活性脂質"尾崎由基男等(金芳堂). 182 (2002)