癌抑制遺伝子の作用機構

• Project/Area Number: 06283215
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas (A)
• Allocation Type: Single-year Grants
• Research Institution: The University of Tokyo (1998-1999); Osaka University (1994-1997)
• Principal Investigator: 秋山 徹 東京大学, 分子細胞生物学研究所, 教授 (70150745)
• Co-Investigator(Kenkyū-buntansha): 石館 宇夫 東京大学, 分子細胞生物学研究所, 助手 (70301116)
• Project Period (FY): 1994 – 1999
• Project Status: Completed (Fiscal Year 1999)
• Budget Amount: ¥117,000,000 (Direct Cost: ¥117,000,000); Fiscal Year 1999: ¥29,000,000 (Direct Cost: ¥29,000,000); Fiscal Year 1998: ¥23,000,000 (Direct Cost: ¥23,000,000); Fiscal Year 1997: ¥20,000,000 (Direct Cost: ¥20,000,000); Fiscal Year 1996: ¥15,000,000 (Direct Cost: ¥15,000,000); Fiscal Year 1995: ¥15,000,000 (Direct Cost: ¥15,000,000); Fiscal Year 1994: ¥15,000,000 (Direct Cost: ¥15,000,000)
• Keywords: 癌抑制遺伝子 / 大腸癌 / APC / Wntシグナル / β-catenin / TCF / G蛋白質 / Axin / D-Axin / Slimb / βTrCP / ICAT / APCL / SPAL / がん抑制遺伝子 / hDLG / b-catenin / D2 / WT1 / 細胞周期 / 大腸がん / DLG / guanylate kinase / ドメイン / SH3ドメイン / armadillo / G1-S移行 / Wilms腫瘍 / Cyclin-CDR / 細胞同期 / cyclin / CDK / 癌 / リン酸化

Research Abstract

(1)Wntシグナル伝達経路は癌化や形態形成に重要な役割をはたしている。Wntシグナルによりβ-cateninは安定化しTCF/LEFと複合体をつくり標的遺伝子の転写を活性化する。我々は、新規β-catenin結合蛋白質を見出し(ICATと命名)、ICATがβ-cateninとTCFの相互作用を阻害することにより標的遺伝子の転写活性化を阻害することを明らかにした。さらに、ICATはWntシグナルを阻害することによりXenopusの体軸形成を阻害する活性をもつことを明らかにした。(2)APC遺伝子は家族性大腸腺腫症の原因遺伝子で一般の大腸癌の大多数でも変異をおこしている。我々は、APC遺伝子産物のarmadilloリピートに結合する新規蛋白質を見出しAsefと命名した。APCとAsefは、大腸、海馬など在が一致し、β-cateninを含む複合体を形成していると考えられた。Asefは、Dblファミリーに属すRac特異的なguanine nucleotide exchange factor(GEF)で、APCと結合することにより活性化した。さらに、MDCK細胞にAsefを過剰発現するとAPC依存的に形態変化、rufflingを誘導した。APC-Asef複合体はアクチン細胞骨格ネットワーク、細胞形態、細胞運動などの制御に関与している可能性があると考えられた。

Research Products

• [Publications] F. Hamada, et al.: "Negative regulation of Wingless signaling by D-Axin, a Drosophila homologue of Axin"Science. 283. 1739-1742 (1999)
• [Publications] F. Hamada, et al.: "Identification and Characterization of D-APC2, a Novel Drosophila Homologue of the Tumor Suppressor APC"Genes Cells. 4. 465-474 (1999)
• [Publications] T. Yamochi, et al.: "Adenovirus-mediated high expression of BCL-6 in CV-1 cells induces apoptotic cell death accompanied by downregulation of BCL-2 and BCL-XL"Oncogene. 18. 487-494 (1999)
• [Publications] T. Tezuka, et al.: "PSD-95 promotes Fyn-mediated tyrosine phosphorylation of the N-methl-D0aspartate receptor subunit NR2A"Proc. Natil. Acad. Sci. USA. 96. 435-440 (1999)
• [Publications] T. Senda, et al.: "Localization of MCC (mutated in colorectal cancer) in various tissues of mice and its involvement in cell differentiation"J Histochem Cytochem. 47. 1149-1158 (1999)
• [Publications] C.R, Badal, et al.: "Localization of the GAP family SPAL in the Rat Esophagus and Heart"Med. Electron Microsc.. 32. 20-24 (1999)
• [Publications] T. Suzuki, et al.: "Tax oncoprotein of HTLV-1 binds to the human homologue of Drosophila discs large tumor suppressor protein, hDLG, and perturbs its function in cell growth control"Oncogene. 18. 5967-5972 (1999)
• [Publications] M. Takemura, et al.: "Nucleolar protein B23.1 binds to retinoblastoma protein and synergistically stimulates DNA polymerase alpha activity"J. Biochem.. 125. 904-909 (1999)
• [Publications] T. Ishidate et al.: "The APC-hDLG Complex Negatively Regulates Cell Cycle Progression from the G0/G1 to S Phase"Oncogene. 19. 365-372 (2000)
• [Publications] H. Nakagawa, et al.: "EB3, a novel member of the EB1 family preferentially expressed in the central nervous system, binds to a CNS-specific APC homologue"Oncogene. 19. 210-216 (2000)
• [Publications] H. Nakagawa, et al.: "APCL, a central nervous system-specific homolog of adenomatous polyposis coli tumor suppressor, binds to p53-binding protein 2 and translocates it to the perinucleus"Cancer Res.. 60. 101-105 (2000)
• [Publications] T. Senda, et al.: "Expression of APC in the mouse kidney and its colocalization with b-catenin and hDLB"J. Histochem. Cytochem. in press..
• [Publications] K. Kawahara, et al.: "Downregulation of b-catenin by the Colorectal Tumor Suppressor APC Requires Association with Axin and B-catenin"J. Biol. Chem. in press..
• [Publications] T.Nakamura et al.: "Axin,an inhibitor of the Wnt signaling pathway,interacts with β-catenin,GSK-3β and APC and reduces the β-catenin level." Genes Cells. 3. 395-403 (1998)
• [Publications] T.Senda et al.: "Expression and localization of the APC tumor suppressor protein in the nervous system." Neuroscience. 83. 857-866 (1998)
• [Publications] K.Inoue et al.: "Wilms' tumor gene (WT1) competes with differentiation-inducing signal in hematopoietic progenitor cells." Blood. 91. 2969-2976 (1998)
• [Publications] H.Nakagawa et al.: "Identification of a brain-specific APC homologue,APCL,and its interaction with β-catenin." Cancer Res.58. 5176-5181 (1998)
• [Publications] Y.Dabashi et al.: "CDK4,a possible critical regulator of apoptosis in rat pheochromocytoma PC12 cells." Biochem.Biophys.Res.Commun.253. 609-613 (1998)
• [Publications] T.Yamagami et al.: "Suppression of Wilms' tumor gene (WT1) expression induces G2/M arrest in leukemic cells." Leuk.Res.22. 383-384 (1998)
• [Publications] K.Mori et al.: "Identification of brain-specific splicing variants of the hDLG1 gene and altered splicing in neuroblastoma cell lines." J.Hum.Genet.43. 123-127 (1998)
• [Publications] T.Yamochi et al.: "Adenovirus-mediated high expression of BCL-6 in CV-1 cells induces apoptotic cell death accompanied by downregulation of BCL-2 and BCL-XL." Oncogene. 18. 487-494 (1999)
• [Publications] T.Tezuka et al.: "PSD-95 promotes Fyn-mediated tyrosine phosphorylation of the N-methyl-D-aspartate receptor subunit NR2A." Proc.Natl.Acad.Sci.USA96. 435-440 (1999)
• [Publications] F.Hamada et al.: "Negative regulation of Wingless signaling by D-Axin,a Drosophila homologue of Axin." Science. 283. 1739-1742
• [Publications] T.Senda et al.: "Association of the MCC (Mutated in colorectal cancer) gene product to the plasma membrane and membrane organelles." Medical Electron Microscopy. (in press).
• [Publications] M.Takemura et al.: "Nucleolar protein B23.1 binds to retinoblastoma protein and synergistically stimulates DNA polymerase alpha activity." J.Biochem.(in press).
• [Publications] R.B.Chandra et al.: "Localization of the GAP family SPAL in the Rat Esophagus and Heart." Medical Electronmicroscopy. (in press).
• [Publications] K.Satoh, et al.: "DAP-1,a novel protein that interacts with the guanylate kinase-like domains of DLG and PSD-95." Genes to Cells. 2. 415-424 (1997)
• [Publications] T.Ishidate, et al.: "Identification of a novel nuclear localization signal in Sam68." FEBS LETTERS. 409. 237-241 (1997)
• [Publications] M.Moriyama, et al.: "The BCL-6 protein is phosphorylated at multiple sites in its serine-and proline-clustered region by mitogen activated protein kinase(MAPK)in vivo." Oncogene. 14. 2465-2474 (1997)
• [Publications] T,Kiyono, et al.: "Binding of high risk HPV E6 oncoproteins to PDZ-domain-containing proteins." Proc.Natl.Acad.Sci.USA. 94. 11616-11612 (1997)
• [Publications] Y.Murata, et al.: "The Wilms tumor suppressor gene WT1 induces G1 arrest and apoptosis in myeloblastic leukemia M1 cells." FEBS LLETTERS.409. 41-45 (1997)
• [Publications] H.Shibata, et al.: "Rapid colorectal adenoma formation initiated by conditional targeting of the Apc gene." Science. 278. 120-123 (1997)
• [Publications] M.Takemura, et al.: "Phosphorylated retinoblastoma protein stimulates DNA polymerase" Oncogene. 15. 2483-2492 (1997)
• [Publications] Y.Dobashi, et al.: "The Wilms tumor protein is persistently associated with the nuclear matrix throughout the cell cycle." Mol.Cell.Biochem.171. 121-126 (1997)
• [Publications] I.Miyashiro, et al.: "Midkine expression in human breast cancers: Expression of truncated form." Breast Can.Res.Treatment. 43. 1-6 (1997)
• [Publications] K.Kagawa, et al.: "Overexpression of GML promotes radiation-induced cell cycle arrest and apoptosis." Biochem.Biophys.Res.Commun.241. 481-485 (1997)
• [Publications] Y.Dobashi, et al.: "Cell cycle control with minimal participation of CDK2 in a murine fibrosarcoma clone cultured in protein-free medium." Biochem.Biophys.Res.Commun.232. 622-625 (1997)
• [Publications] J.Adachi, et al.: "Growth suppression of non-small cell lung carcinoma cells by the introduction of the p16INK4A gene." Int J.Cancer. 10. 33-39
• [Publications] T.Senda, et al.: "Expression and localization of the APC tumor suppressor protein in the nervous system." Neuroscience. 83. 857-866 (1998)
• [Publications] T.Yamagami, et al.: "Suppression of Wilms tumor gene(WT1)expression induces G2/M arest in leukemic cells." Leukemia Res.(in press).
• [Publications] T.Senda, et al.: "Association of the MCC(Mutated in colorectal cancer)gene product to the plasma membrane and membrane organelles." Medical Electron Microscopy in press.
• [Publications] K.Inoue, et al.: "Wilms'tumor gene(WT1) competes with defferentiation-induced signal in hematopoietic progenitor cells." Blood. (in press).
• [Publications] A.Matsumine,et al.: "The MCC gene product regulates cell cycle transition from G0/G1 to S phase." J.Biol.Chem.271. 10341-10346 (1996)
• [Publications] A.Matsumine,et al.: "Binding of APC to the human homolog of the Drosophila Discs large tumor suppressor protein." Science. 272. 1020-1023 (1996)
• [Publications] T.Kudoh,et al.: "Constitutive expression of the Wilms tumor suppressor gene WT1 in F9 embryonal carcinoma cells induces apoptotic cell death in response to retinoic acid." Oncogene.13. 1431-1439 (1996)
• [Publications] Y.Adachi,et al.: "Midkine as a novel target gene for the Wilms'tumor suppressor gene(WT1)." Oncogene. 13. 2203-2197 (1996)
• [Publications] T.Yamagami,et al.: "Growth inhibition of human leukemic cells by WT1(Wilms tumor gene)antisense oligodeoxynucleotides:implications for the involvement of WT1 in leukemogenesis." Blood. 87. 2878-2884 (1996)
• [Publications] R.N.Bhattacharjee,et al.: "The tumor suppressor gene product APC is hyperphosphorylated during the M phase." Biochem.Biophys.Res.Commun.220. 192-195
• [Publications] 秋山徹: "「がん 最新知見から病態をとらえる」" メジカルビュー社, 160 (1997)
• [Publications] 秋山徹: "「医科分子生物学 改定第3版」" 南江堂, 501 (1997)
• [Publications] T.Kudou et al.: "G1 phase arrest induced by Wilms tumor protein WT1 is abrogated by cyclin/CDK." Proc.Natl.Acad.Sci.USA.92. 4517-4521 (1995)
• [Publications] I.Miyashiro et al.: "Subcellular localization of the APC protein:Immunoelectron microscopic study of the association of the APC protein with catenin." Oncogene.11. 89-96 (1995)
• [Publications] Y.Dobashi et al.: "Constitutive overexpression of Cdk2 inhibits neuronal differentiation of rat pheochromocytoma PC12 cells." J.Biol.Chem.270. 23031-23037 (1995)
• [Publications] G-H,Baeg, et al.: "The tumor suppressor gene product APC blocks cell cycle progression from the G0/G1 to S phase." EMBOJ. 14. 5618-5625 (1995)
• [Publications] A.Matsumine,et al.: "The MCC gene product regulates cell cycle transition from G0/G1 to S phase." J.Biol.Chem.(in press).
• [Publications] T.Yamagami,et al.: "Growth inhibition of human leukemic cells by WT1 antisense oligodeoxynucleotides:implications for the involvement of WT1 in leukemogenesis" Blood.(in press).
• [Publications] 秋山 徹: "癌抑制遺伝子の最前線" 横田 淳 秋山 徹, 158 (1995)
• [Publications] 秋山 徹: "転写のしくみと疾患" 村松 正寛, 163 (1996)
• [Publications] S.Kamada et al.: "Serum-independent phosphory lation of C-Jum and alteration in AP-1 componets by transformation with variousoncogenes" J.Biol.Chem.269. 4565-4570 (1994)
• [Publications] K.Inoue et al.: "WT1 as a new proguostic factor and a new marker for the detection of minimal residual disease in acute leukemia" Blood. 84. 3071-3079 (1994)
• [Publications] T.Kudoh et al.: "GI phase arrest induced by Wilmstumor protein WTI is abrogated by Cyclin ICDK" Proc.Natl.Acad,Sci,USA. 92(inpress). (1995)
• [Publications] 豊島久真男・秋山徹: "癌化のシグナル、伝達機構" 中外医学社, 294 (1994)
• [Publications] 豊島久真男他: "がんと遺伝子" 化学同人, 149 (1994)