細胞がん化のシグナル伝達の解析と抗がん剤リードの探索

• Project/Area Number: 12217163
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: National Institute of Infectious Diseases
• Principal Investigator: 上原 至雅 国立感染症研究所, 生物活性物質部, 部長 (50160213)
• Co-Investigator(Kenkyū-buntansha): 深澤 秀輔 国立感染症研究所, 生物活性物質部, 室長 (10218878) ; 村上 裕子 国立感染症研究所, 生物活性物質部, 主任研究官 (00142133) ; 野口 耕司 国立感染症研究所, 生物活性物質部, 主任研究官 (80291136)
• Project Period (FY): 2000 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥115,500,000 (Direct Cost: ¥115,500,000); Fiscal Year 2004: ¥22,500,000 (Direct Cost: ¥22,500,000); Fiscal Year 2003: ¥22,300,000 (Direct Cost: ¥22,300,000); Fiscal Year 2002: ¥22,000,000 (Direct Cost: ¥22,000,000); Fiscal Year 2001: ¥24,700,000 (Direct Cost: ¥24,700,000); Fiscal Year 2000: ¥24,000,000 (Direct Cost: ¥24,000,000)
• Keywords: 足場非依存性増殖 / アノイーキス / アポトーシス / BH3-only蛋白質 / Bim / シグナル伝達 / MEK阻害剤 / apoptosis / anoikis / 乳がん / ERK / anicequol / spicamycin / 乳がん細胞 / U0126 / rapamycin / MIA PaCa-2 / TPA

Research Abstract

MEK阻害剤が、ヒト乳がん細胞2株に対し、非接着状態でのみapoptosis、即ちanoikis感受性を誘導すること、その感受性誘導にBH3-only proteinの一つであるBimELが重要な働きをしていることを報告した。BimELはERKによるリン酸化がユビキチン化の引き金になっており、MEK阻害剤はそのリン酸化を阻害する。その結果ユビキチン-プロテオソーム系によるBimELの分解が抑制され、BimELの蛋白レベルが上昇する。anoikis感受性が誘導されるがん細胞株では、もともとのBimの蛋白レベルが非常に低く、これらの細胞ではMEK-ERK経路の活性化によるBimELのリン酸化、分解促進がanoikis回避の主要な機構であると考えられた。乳がん以外のがん細胞でもMEK阻害剤によりanoikis感受性となるものがあり、特に大腸がん細胞株は程度の差はあるものの、調べた株全てでanoikis感受性が誘導された。いずれの細胞でもBimELレベルは上昇し、anoikis感受性の誘導はERKの活性化レベルよりも、BimELの増加と相関した。以上から、Bimはanoikis誘導の重要な決定因子であり、MEK阻害剤感受性予測の指標となることが示唆された。また、がん化シグナル特異的阻害剤の探索からは、放線菌よりspicamycinの新規類縁体を見出す一方、真菌由来のテルペノイドanicequolの誘導体の中に、anoikis誘導活性においてanicequolより選択性が大きい化合物を見出した。

Research Products

• [Journal Article] BimEL is an important determinant for induction of anoikis sensitivity by mitogen-activated protein/extracellular signal-regulated kinase kinase (MEK) inhibitors. (2004)
  • Author(s): Fukazawa, H.
  • Journal Title: Mol.Cancer Therap. 3 Pages: 1281-1288
• [Journal Article] Nucleolar Nek11 is a novel target of Nek2A in G1/S-arrested cells. (2004)
  • Author(s): Noguchi, K.
  • Journal Title: J.Biol.Chem. 279 Pages: 32716-32727
• [Journal Article] Synthesis and biological evaluation of benzamindes and benzamidines : structural requirement of a pyrimidine ring for inhibition of EGFR tyrosine kinase. (2004)
  • Author(s): Asano, T.
  • Journal Title: Bioorg.Med.Chem.Lett. 14 Pages: 2299-2302
• [Journal Article] Design, synthesis, and biological evaluation of aminoboronic acids as growth-factor receptor inhibitors for EGFR and VEGFR-1 yrosine kinases. (2004)
  • Author(s): Asano, T.
  • Journal Title: Chembiochem. 5 Pages: 483-490
• [Journal Article] Benzamides and benzamidines as specific inhibitors of epidermal growth factor receptor and v-Src protein tyrosine kinases. (2004)
  • Author(s): Asano, T.
  • Journal Title: Bioorg.Med.Chem. 12 Pages: 3529-3542
• [Journal Article] Poly(ADP-ribose) polymerase-1 inhibits ATM kinase activity in DNA damage response. (2004)
  • Author(s): Watanabe, F.
  • Journal Title: Biochem.Biophys.Res.Commun. 319 Pages: 596-602
• [Journal Article] Destruxin E, a cyclodepsipeptide antibiotic, reduces cyclin D1 levels and inhibits anchorage-independent growth of v-Ki-ras-expressed pMAM-ras-REF cells. (2004)
  • Author(s): Kobayashi, T.
  • Journal Title: Biol.Pharm.Bull. 27 Pages: 587-590
• [Journal Article] Design of hypoxia targeting protein tyrosine kinase inhibitor using an innovative pharmacophore 2-methylene-4-cyclopentene-1,3-dione. (2004)
  • Author(s): Hori, H.
  • Journal Title: Biochim.Biophys.Acta 1697 Pages: 29-38
• [Patent(Industrial Property Rights)] 抗ガン作用を有する化合物、その製造方法及び抗ガン剤 (2004)
  • Inventor(s): 上原 至雅, 深澤 秀輔, 古米 保, 五十嵐 康弘
  • Industrial Property Rights Holder: ヒューマンサイエンス振興財団
  • Industrial Property Number: 2004-186572
  • Filing Date: 2004-06-24
• [Publications] Gutzkow, KB.: "Cyclic AMP inhibits translation of cyclin D3 in T-lymphocytes at the level of elongation by inducing eEF2-phosphorylation"Cellular Signalling. 15. 871-881 (2003)
• [Publications] Uehara, Y.: "Natural product origins of Hsp90 inhibitors."Current Cancer Drug Targets. 3. 319-324 (2003)
• [Publications] Hori, H.: "Design of hypoxia-targeting protein tyrosine kinase inhibitor using an innovative pharmacophore 2-methylene-4-cyclopentene-1,3-dione"Biochim.Biophys.Acta. 1697. 29-38 (2003)
• [Publications] Kamei, K.: "The trend of imported mycoses in Japan"J.Infec.Chemother.. 9. 16-20 (2003)
• [Publications] 上原 至雅: "プロテインキナーゼ阻害薬"がん分子標的治療. 1-3. 18-25 (2003)
• [Publications] Fukazawa, H: "Mitogen-activated protein/extracellular signal-regulated kinase kinase (MEK) inhibitors restore anoikis sensitivity in human breast cancer cell lines with a constitutively activated extracellular-regulated kinase (ERK) pathway"Mol.Cancer Ther.. 1. 303-309 (2002)
• [Publications] Noguchi, K.: "Nekll, a new member of the NIMA family of kinases, involved in DNA replication and genotoxic stress responses"J.Biol.Chem.. 277. 39655-39665 (2002)
• [Publications] Ishino, K.: "Enhancement of anchorage-independent growth of human pancreatic carcinoma MIA PaCa-2 cells by signaling from protein kinase C to mitogen-activated protein kinase"Mol.Carcinog.. 34. 180-186 (2002)
• [Publications] Cho, S.I.: "Effects of hibarirnicins and hibarimicin-related compounds produced by Microbispora on v-Src kinase activity and growth and differentiation of human myeloid leukemia HL-60 cells"J.Antibiot.. 55. 270-278 (2002)
• [Publications] Hori, H.: "TX-1123 : an antitumor 2-hydroxyarylidene-4-cyclopentene-1, 3-dione as a protein tyrosine kinase inhibitor having low mitochondrial toxicity"Bioorg.Med.Chem.. 10. 3257-3265 (2002)
• [Publications] Murakami, Y.: "A mammalian two-hybrid screening system for inhibitors of interaction between HIV Nef and the cellular tyrosine kinase Hck"Antiviral.Res.. 55. (2002)
• [Publications] Wada, S.: "Candida glabrata ATP-binding cassette transporters Cdrlp and Pdhlp expressed in a Saccharomyces cerevisiae strain deficient in membrane transporters show phosphorylation-dependent pumping properties"J.Biol.Chem.. 277. 46809-46821 (2002)
• [Publications] Umeyama, T.: "Construction of FLAG tagging vectors for Candida albicans"Yeast. 19. 611-618 (2002)
• [Publications] Igarashi, Y.: "Anicequol, a novel inhibitor for anchorage-independent growth of tumor cells from Penicillium aurantiogriseum Dierckx TP-F0213"J.Antibiot.. 55. 371-376 (2002)
• [Publications] Hori, H.: "Biosynthesis of hibarimicins.I.13C-labeling experiments"J.Antibiot.. 55. 46-52 (2002)
• [Publications] Kajiura, T.: "Biosynthesis of hibarimicins. II. Elucidation of biosynthetic pathway by cosynthesis using blocked mutants"J.Antibiot.. 55. 53-60 (2002)
• [Publications] Igarashi, Y.: "Biosynthesis of hibarimicins. III. Structures of new hibarimicin-related metabolites produced by blocked mutants"J.Antibiot.. 55. 61-70 (2002)
• [Publications] Yamada, T.: "Leptosins M-N1, cytotoxic metabolites from Leptosphearia species separated from a marine alga. Structure determination and biological activities"Tetrahedron. 58. 479-487 (2002)
• [Publications] Hashimoto, S.: "Beta-Hydroxyisovaleryishikonin is a novel and potent inhibitor of protein tyrosine kinases"Jpn.J.Cancer Res.. 93. 944-951 (2002)
• [Publications] Noguchi, K.: "ASK1-signaling promotes c-Myc protein stability during apoptosis"Biochem Biophys Res Commun.. 281. 1313-1320 (2002)
• [Publications] Hori, H.: "Biosynthesis of hibarimicins. I. ^<13>C-Labeling experiments"J.Antibiotics. 55. 46-52 (2002)
• [Publications] Kajiura, T.: "Biosynthesis of hibarimicins. II. Elucidation of biosynthetic pathway by cosynthesis using blocked mutants"J.Antibiotics. 55. 53-60 (2002)
• [Publications] Igarashi, Y.: "Biosynthesis of hibarimicins. III. Structures of new hibarimicin-related metabolites produced by blocked mutants"J.Antibiotics. 55. 61-70 (2002)
• [Publications] Cho, S.I.: "Effects of hibarimicins and hibarimicin-related compounds produced by Microbispora on v-src kinase activity and growth and differentiation of human myeloid leukemia HL-60 cells"J.Antibiotics. (in press).
• [Publications] Murakami, Y.: "A mammalian two-hybrid screening system for inhibitors of interaction between HIV Nef and the cellular tyrosine kinase Hck"Antiviral Res.. (in press).
• [Publications] Kitanaka, C.: "Increased Ras expression and caspase-independent neuroblastoma cell death : possible mechanism of spontaneous neuroblastoma regression"J.Natl.Cancer Inst.. (in press).
• [Publications] Fukazawa, H.: "Mitogen-activated Protein/Extracellular Signal-regulated Kinase Kinase(MEK)inhibitors restore anoikis-sensitivity in human breast cancer cell lines with constitutively activated Extracellular Signal-regulated Kinase(ERK)pathway"Mol. Cancer Therapeut. (in press).
• [Publications] Igarashi, Y.: "Anicequol, a novel inhibitor for anchorage-independent growth of tumor cells from Penicillium sp. TP-F0213"J.Antibiotics. (in press).
• [Publications] Endo,N.: "Herbimycin A induces G1 arrest through accumulation of p27Kip1 in cyclin D1-overexpressing fibrobliasts."Biochem.Biophys.Res.Commun.. 267. 54-58 (2000)
• [Publications] Fukazawa,H.: "U0126 reverses Ki-ras mediated transformation by blocking both MAP kinase and p70 S6 kinase pathways."Cancer Res.. 60. 2104-2107 (2000)
• [Publications] Cho,S.I.: "Novel compounds,'1,3-selenazine derivatives' as specific inhibitors of eukaryotic elongation factor 2-kinase."Biochem.Biophys.Acta. 1475. 207-215 (2000)