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

• 研究課題/領域番号: 12217163
• 研究種目: 特定領域研究
• 配分区分: 補助金
• 審査区分: 生物系
• 研究機関: 国立感染症研究所
• 研究代表者: 上原 至雅 国立感染症研究所, 生物活性物質部, 部長 (50160213)
• 研究分担者: 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)
• 研究期間 (年度): 2000 – 2004
• 研究課題ステータス: 完了 (2004年度)
• 配分額: 115,500千円 (直接経費: 115,500千円); 2004年度: 22,500千円 (直接経費: 22,500千円); 2003年度: 22,300千円 (直接経費: 22,300千円); 2002年度: 22,000千円 (直接経費: 22,000千円); 2001年度: 24,700千円 (直接経費: 24,700千円); 2000年度: 24,000千円 (直接経費: 24,000千円)
• キーワード: 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

研究概要

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。

研究成果

• [雑誌論文] BimEL is an important determinant for induction of anoikis sensitivity by mitogen-activated protein/extracellular signal-regulated kinase kinase (MEK) inhibitors. (2004)
  • 著者名/発表者名: Fukazawa, H.
  • 雑誌名: Mol.Cancer Therap. 3 ページ: 1281-1288
• [雑誌論文] Nucleolar Nek11 is a novel target of Nek2A in G1/S-arrested cells. (2004)
  • 著者名/発表者名: Noguchi, K.
  • 雑誌名: J.Biol.Chem. 279 ページ: 32716-32727
• [雑誌論文] Synthesis and biological evaluation of benzamindes and benzamidines : structural requirement of a pyrimidine ring for inhibition of EGFR tyrosine kinase. (2004)
  • 著者名/発表者名: Asano, T.
  • 雑誌名: Bioorg.Med.Chem.Lett. 14 ページ: 2299-2302
• [雑誌論文] Design, synthesis, and biological evaluation of aminoboronic acids as growth-factor receptor inhibitors for EGFR and VEGFR-1 yrosine kinases. (2004)
  • 著者名/発表者名: Asano, T.
  • 雑誌名: Chembiochem. 5 ページ: 483-490
• [雑誌論文] Benzamides and benzamidines as specific inhibitors of epidermal growth factor receptor and v-Src protein tyrosine kinases. (2004)
  • 著者名/発表者名: Asano, T.
  • 雑誌名: Bioorg.Med.Chem. 12 ページ: 3529-3542
• [雑誌論文] Poly(ADP-ribose) polymerase-1 inhibits ATM kinase activity in DNA damage response. (2004)
  • 著者名/発表者名: Watanabe, F.
  • 雑誌名: Biochem.Biophys.Res.Commun. 319 ページ: 596-602
• [雑誌論文] Destruxin E, a cyclodepsipeptide antibiotic, reduces cyclin D1 levels and inhibits anchorage-independent growth of v-Ki-ras-expressed pMAM-ras-REF cells. (2004)
  • 著者名/発表者名: Kobayashi, T.
  • 雑誌名: Biol.Pharm.Bull. 27 ページ: 587-590
• [雑誌論文] Design of hypoxia targeting protein tyrosine kinase inhibitor using an innovative pharmacophore 2-methylene-4-cyclopentene-1,3-dione. (2004)
  • 著者名/発表者名: Hori, H.
  • 雑誌名: Biochim.Biophys.Acta 1697 ページ: 29-38
• [産業財産権] 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)
  • 発明者名: 上原 至雅, 深澤 秀輔, 古米 保, 五十嵐 康弘
  • 権利者名: ヒューマンサイエンス振興財団
  • 産業財産権番号: 2004-186572
  • 出願年月日: 2004-06-24
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• [文献書誌] Noguchi, K.: "ASK1-signaling promotes c-Myc protein stability during apoptosis"Biochem Biophys Res Commun.. 281. 1313-1320 (2002)
• [文献書誌] Hori, H.: "Biosynthesis of hibarimicins. I. ^<13>C-Labeling experiments"J.Antibiotics. 55. 46-52 (2002)
• [文献書誌] Kajiura, T.: "Biosynthesis of hibarimicins. II. Elucidation of biosynthetic pathway by cosynthesis using blocked mutants"J.Antibiotics. 55. 53-60 (2002)
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• [文献書誌] 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).
• [文献書誌] Kitanaka, C.: "Increased Ras expression and caspase-independent neuroblastoma cell death : possible mechanism of spontaneous neuroblastoma regression"J.Natl.Cancer Inst.. (in press).
• [文献書誌] 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).
• [文献書誌] Igarashi, Y.: "Anicequol, a novel inhibitor for anchorage-independent growth of tumor cells from Penicillium sp. TP-F0213"J.Antibiotics. (in press).
• [文献書誌] Endo,N.: "Herbimycin A induces G1 arrest through accumulation of p27Kip1 in cyclin D1-overexpressing fibrobliasts."Biochem.Biophys.Res.Commun.. 267. 54-58 (2000)
• [文献書誌] Fukazawa,H.: "U0126 reverses Ki-ras mediated transformation by blocking both MAP kinase and p70 S6 kinase pathways."Cancer Res.. 60. 2104-2107 (2000)
• [文献書誌] 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)