2002 年度実績報告書

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

研究課題/領域番号	12217163
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	村上裕子国立感染研究所, 生物活性物質部, 主任研究官 (00142133) 深澤秀輔国立感染研究所, 生物活性物質部, 室長 (10218878)
キーワード	MEK阻害剤 / anicequol / 足場非依存性増殖 / apoptosis / 乳がん細胞 / anoikis / Bim / ERK
研究概要	我々は昨年度、MEK阻害剤が、ERKが恒常的に活性化しているヒト乳がん細胞に対しanoikis感受性を誘導することを報告した。今年度はMEK阻害剤によるanoikis感受性誘導を規定する因子を明らかにするため、apoptosis関連蛋白質に対する影響を調べ、MEK阻害剤処理によりBH3-onlyproteinの一つであるBimELの蛋白量が増加することを見いだした。電気泳動上の移動度の変化から、MEK阻害剤処理細胞ではBimELのリン酸化が低下していることが予想された。in vitroでBimELはERKによりリン酸化されたので、そのリン酸化部位を決定した。この残基は実際に細胞内でもERKによりリン酸化されることが示唆された。細胞によってはERKにょるリン酸化がBim蛋白質の減少を引き起こし、anoikis回避に重要な役割を果たしているものと思われる。anoikis感受性にならない細胞では、BimELの増加は観察されず、BimELはanoikis感受性誘導の重要な決定要因であると考えられた。昨年に続き、糸状菌由来テルペノイドanicequolのがん細胞足場非依存的増殖阻害作用機構を解析した。直接の標的を同定するため、ビオチン化化合物を作製し、結合蛋白質をいくつか検出した。またがん細胞の足場非依存性増殖阻害を指標に海産物、放線菌、真菌に抗がん剤シーズを探索し、新たに3物質を同定した。

研究成果

(14件)

すべてその他

すべて文献書誌 (14件)

[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 a constitutively activated extracellular-regulated kinase (ERK) pathway"Mol.Cancer Ther.. 1. 303-309 (2002)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[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.. 55. (2002)
[文献書誌] 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)
[文献書誌] Umeyama, T.: "Construction of FLAG tagging vectors for Candida albicans"Yeast. 19. 611-618 (2002)
[文献書誌] 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)
[文献書誌] Hori, H.: "Biosynthesis of hibarimicins.I.13C-labeling experiments"J.Antibiot.. 55. 46-52 (2002)
[文献書誌] Kajiura, T.: "Biosynthesis of hibarimicins. II. Elucidation of biosynthetic pathway by cosynthesis using blocked mutants"J.Antibiot.. 55. 53-60 (2002)
[文献書誌] Igarashi, Y.: "Biosynthesis of hibarimicins. III. Structures of new hibarimicin-related metabolites produced by blocked mutants"J.Antibiot.. 55. 61-70 (2002)
[文献書誌] 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)
[文献書誌] Hashimoto, S.: "Beta-Hydroxyisovaleryishikonin is a novel and potent inhibitor of protein tyrosine kinases"Jpn.J.Cancer Res.. 93. 944-951 (2002)

2002 年度 実績報告書

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)

研究成果

[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 a constitutively activated extracellular-regulated kinase (ERK) pathway"Mol.Cancer Ther.. 1. 303-309 (2002)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[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.. 55. (2002)

[文献書誌] 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)

[文献書誌] Umeyama, T.: "Construction of FLAG tagging vectors for Candida albicans"Yeast. 19. 611-618 (2002)

[文献書誌] 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)

[文献書誌] Hori, H.: "Biosynthesis of hibarimicins.I.13C-labeling experiments"J.Antibiot.. 55. 46-52 (2002)

[文献書誌] Kajiura, T.: "Biosynthesis of hibarimicins. II. Elucidation of biosynthetic pathway by cosynthesis using blocked mutants"J.Antibiot.. 55. 53-60 (2002)

[文献書誌] Igarashi, Y.: "Biosynthesis of hibarimicins. III. Structures of new hibarimicin-related metabolites produced by blocked mutants"J.Antibiot.. 55. 61-70 (2002)

[文献書誌] 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)

[文献書誌] Hashimoto, S.: "Beta-Hydroxyisovaleryishikonin is a novel and potent inhibitor of protein tyrosine kinases"Jpn.J.Cancer Res.. 93. 944-951 (2002)

2002 年度実績報告書

上原至雅国立感染症研究所, 生物活性物質部, 部長 (50160213)