がんの発生、発達に関する情報伝達の解析

• Project/Area Number: 10152101
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas (A)
• Allocation Type: Single-year Grants
• Research Institution: The University of Tokyo
• Principal Investigator: 福井 泰久 東京大学, 大学院・農学生命科学研究科, 教授 (00181248)
• Co-Investigator(Kenkyū-buntansha): 黒崎 知博 関西医科大学, 医学部, 教授 (50178125) ; 稲垣 昌樹 愛知県がんセンター研究所, 生化学部, 部長(研究職) (30183007) ; 一條 秀憲 東京医科歯科大学, 歯学部, 教授 (00242206) ; 田村 真理 東北大学, 加齢医学研究所, 教授 (20124604) ; 白井 隆一 奈良先端大学, 大学院・物質科学教育研究センター, 助教授 (80183838)
• Project Period (FY): 1999
• Project Status: Completed (Fiscal Year 1999)
• Budget Amount: ¥74,000,000 (Direct Cost: ¥74,000,000); Fiscal Year 1999: ¥37,000,000 (Direct Cost: ¥37,000,000); Fiscal Year 1998: ¥37,000,000 (Direct Cost: ¥37,000,000)
• Keywords: リン脂質 / ホシファテシジルイノシトールキナーゼ / 組織応答 / 分化 / アダプター / イノシトールリン脂質 / MAPキナーゼ / ホスホリパーゼ / がん / 情報伝達 / キナーゼ / Gタンパク質 / 細胞応答 / 発癌 / 転移

Research Abstract

膜の構成成分であるリン脂質をはじめとする脂質が細胞の分化・増殖の調節に関わっていることは最近の研究から明らかにされつつある。リン脂質の代謝酵素PI3キナーゼは多機能2次メッセンジャーPIP_3を生じる。また、PLCはDAGおよびIP_3という重要な2次メッセンジャーを生じる。これらの下流には細胞骨格系や種々のキナーゼを介した情報伝達系の調節が位置している。従って、これらの役割を明らかにしていけば、がん細胞の成り立ちを解析でき、がんに対抗するための手段を見いだせると考えられる。本研究は8人の班員がそれぞれお互いに協力しつつ、以下のような成果を挙げた。
1)印環細胞がんの発生にはPI3キナーゼが関与しており、その下流因子p38が重要な役割をしていること、その阻害剤が印環細胞がん抑制に有効な可能性を示した。
2)PIP_3の2位は天然には不飽和脂肪酸が占めているが、飽和脂肪酸でも短鎖であれば生活性を持つことを示し、今後のPIP_3アナログ作成を容易にした。
3)PLCに関してδ4の調節機構とその生理的役割を明らかにするとともに、新規タンパク質L2を同定し、その役割について検討した。
4)PKCηのノックアウトマウスを作成し、PKCηが発がんプロモーションの抑制に関与していることを明らかにした。
5)BLNKはPLC-γ2と結合し、活性化するが、cb1はこれと拮抗することにより、PLC-γ2を負に調節していることを明らかにした。
6)PPC2一β1はMKKKの一種であるTAK1シグナル伝達を負に制御することを明らかにした。
7)酵母two-hybrid法を用いて、ビメンチン、GFAP、デスミン、およびケラチン18と結合する蛋白質を探索し、いくつかの候補タンパク質を得た。
8)ASK1のTNF-TRAF2系による活性化機構を明らかにするとともに、ASK1の下流因子JNK、p38の細胞応答における役割を明らかにした。

Research Products

• [Publications] Fukui, Y. et al.: "Dedifferentiation of Adenocarcinomas by Activation of Phosphatidylinositol 3-kinase"Proc Natl Acad Sci USA. 96. 4874-4879 (1999)
• [Publications] Fukui, Y. et al.: "The structural requirement of phosphatidylinositols as substrate of phosphatidylinositol 3-kinase"Tetrahed. Let.. 40. 1693-1696 (1999)
• [Publications] Fukui, Y. et al.: "Cyclin Dl expression mediated by phosphatidylinositol 3-kinase through mTOR-p70S6K-independent signaling in growth factor-stimulated NH3T3 fibroblasts."Mol. Cell. Biol.. 19. 1346-1358 (1999)
• [Publications] Fukui, Y. et al.: "Increased insulin sensitivity and hypoglycemia in mice lacking p85a regulatory subunit of phosphoinositide 3-kinase."Nat. Genet.. 21. 230-235 (1999)
• [Publications] Fukui, Y. et al.: "An evidence that a phosphatidylinositol 3,4,5-trisphosphate binding protein can function in nucleus."J. Biol Chem.. 274. 3919-3922 (1999)
• [Publications] Fukui, Y. et al.: "Identification of protein kinase B(PKB) as a phosphatidylinositol 3,4,5-trisphosphate binding protein in Dictyostelium discoideum."Biosci. Biotech. Biochem.. 63. 368-372 (1999)
• [Publications] Fukui, Y. et al.: "Adenoviral gene transfer of activated phosphatidylinositol 3'-kinase and Akt inhibits apoptosis of hypoxic cardimyocytes in vitro"Circulation. 100. 2373-2379 (1999)
• [Publications] Fukui, Y. et al.: "Dedifferentiation of adenocarcinomas by activation of phosphatidylinositol 3-kinase"Proc Natl Cad Sci USA. 96. 4874-4879 (1999)
• [Publications] Fukui, Y. et al.: "In vitro and in vivo suppression of osteoclast function by adenovirus vector-induced csk gene"J Bone Miner Res.. 15. 41-51 (2000)
• [Publications] Chida, K., Fukui. Y., et al.: "How signal transduction systems look like after recent dramatic progress"Tanpakushitsu Kakusan Koso. 100. 2373-2379 (1999)
• [Publications] Chida, K., et al.: "Nuclear microinjection of EDTA inhibits transient expression of the lac-Z gene and stable transformation by the neomycin-resistant gene in NRK cells possibly by causing disruption of nuclear pore function. Circulation"Tissue Cult. Res. Commun.. 18. 119-123 (1999)
• [Publications] Fukami, K., et al.: "A novel phospholipase Cδ4 splice variant as a negative regulator of PLC>"Biol. Chem.. 274. 2872-2879 (1999)
• [Publications] Fukami, K., et al.: "Growth factor-induced promoter activation of murine phospholipase C delta 4."Eur. J. Biochem.. 267. 28-36 (2000)
• [Publications] Tamura, S., et al.: "Proteinphosphatase 2C inactivates F-action binding of human platelet moesin."J. Biol. Chem.. 274. 26705-26712 (1999)
• [Publications] Shirai, R., et al.: "The structural requirement of phosphatidylinositols as substrate of phosphatidylinositol 3- kinase."Tetrahedron Lett.. 40. 1693-1696 (1999)
• [Publications] Ichijo, H., et al.: "Roles of bone morphogenetic protein type I receptors and Smad proteins in osteoblastic and chondroblastic differentiation"Mol. Biol. Cell. 10. 3801-3813 (1999)
• [Publications] Ichijo, H., et al.: "Bcl-2 is phosphorylated and inactivated by an ASK1-JNK pathway morally activated at G2/M"Biol. Cell.. 10. 8469-8478 (1999)
• [Publications] Ichijo, H., et al.: "Induction of apoptosis signal regulation kinase 1 (ASK1) after spinal cord injury in rats: Possible involvement of ASK1-JNK and -p38 pathways in neuronal apoptosis. Circulation"J. Neuropathol. Exp. Neurol.. 58. 442-450 (1999)
• [Publications] Ichijo, H., et al.: "Ask mediates apoptosis cell death induced by geotaxis stress. Circulation"Oncogene. 18. 173-180 (1999)
• [Publications] Ichijo, H., et al.: "Microtuble disfunction induced by Paclitaxel initiates apoptosis through both c-Jun N-terminal kinase (JNK)-dependent and -independent pathways in ovarian cancer cells. Circulation"J. Biol. Chem.. 274. 8208-8216 (1999)
• [Publications] Ichijo, H., et al.: "Apoptosis inhibitory activity of cytoplasmic p21Cipl/WAF1 in monocytic differentiation"EMBO J.. 18. 1223-1234 (1999)
• [Publications] Ichijo, H., et al.: "Triggering neuronal cell death by the in situ activation f SEK1 nuclear polyglutamine aggregations"Oncogene. 18. 2783-2788 (1999)
• [Publications] Inagaki, M., et al.: "Identification of a novel phosphorylation site on histone H3 coupled with mitotic chromosome condensation"J. Biol. Chem.. 274. 25543-25549 (1999)
• [Publications] Inagaki, M., et al.: "Balance between activities of Rho-kinase and Type I protein Phosphatase modulates turnover of phosphorylation and dynamics of desmin/vimentin firmaments"J. Biol. Chem.. 274. 34932-34939 (1999)
• [Publications] Tamura. S., et al.: "Alternative promoters direct tissue-specific expression of the mouse protein phosphatase 2C β gene."Eur. J. Biochem.. 263. 736-745 (1999)
• [Publications] Tamura. S., et al.: "Expressions of four major protein ser/thr phosphatases in human primary leukemic cells"Leukemia. 13. 595-600 (1999)
• [Publications] Kurosaki, T., et al.: "Paired Immunoglobulin-like receptor B (PIR-B) inhibits BCR-induced activation of Syk and Btk by SHP-1. Circulation"Oncogene. 18. 2291-2297 (1999)
• [Publications] Kurosaki, T., et al.: "BLMK required for coupling Syk to PLC-γ2 and Racl-JNK in B cells"Immunity. 10. 117-125 (1999)
• [Publications] Kurosaki, T., et al.: "Encoding Ca^<2+> signals by differential expression of IP_3 receptor subtypes"EMBO J.. 18. 1303-1308 (1999)
• [Publications] Kurosaki, T., et al.: "Inhibitory modulation of B cell receptor-mediated CA^<2+> mobilization by Srchomology 2 domain-containing inositol 5'-phosphatase (SHIP)"J. Biol. Chem..
• [Publications] Kurosaki, T., et al.: "Bruton's tyrosine kinase activity in negatively regulated by Sab, the Btk-SH3 domain binding protein"Proc. Natl. Acad. Sci. U.S.A.. 96. 6341-6346 (1999)
• [Publications] Kurosaki, T., et al.: "In vitro and in vivo suppression of osteoclast function by adenovirus vector-induced csk gene"J Bone Miner Res.. 15. 753-760 (1999)
• [Publications] Kita,Y.et al: "Microinjection of activated phosphatidylinositol-3 kinase induces precess outgrowth in rat PC12 cells through the Tac-JNK signal transd uction pathway." J.Cell.Sci.111. 907-915 (1998)
• [Publications] Shirai,T.et al: "Specific detection of phosphatidylinositol3,4,5-trisphosphate binding proteins by the PIP3 analogus beads:An application for rapid purification of the PIP3 binding proteins" Biochem.Biophys.Acta. 1402. 292-302 (1998)
• [Publications] Fukui,Y.et al.: "Dovnstream of phosphatidylinositol-3 kinase,a multifunctional signaling moleculs,and its regulation in cell responses." J.Biochem.124. 1-7 (1998)
• [Publications] Takawa,N.et al: "Cyclin D1 expression mediated by phosphatidylinositol 3-kinase through mTOR-p70^<S6K>-independent signaling in growth factor-stimulated NIH3T3 fibroblasts." Mol.Cell.Biol.(in press). (1999)
• [Publications] Tanaka,K.et al: "Identification of protein kinase B(PKB)as a phosphatidylinositol3,4,5-trisphosphate binding protein in Dictyostelium discoideum." Biosci.Biotech.Biochem.(in press). (1999)
• [Publications] Tanaka,K.et al: "An evidence that a phosphatidylinositol3,4,5-trisphosphate binding protein can function in nucleus." J.Biol.Chem.(in press). (1999)