2001 Fiscal Year Annual Research Report

アポトーシス情報伝達に着目した新規ヌクレオシド系制癌剤の開発-ネコ白血病、イヌリンパ腫への応用-

Research Project

Project/Area Number	12460135
Research Institution	HOKKAIDO UNIVERSITY
Principal Investigator	桑原幹典北海道大学, 大学院・獣医学研究科, 教授 (10002081)
Co-Investigator(Kenkyū-buntansha)	太田利男北海道大学, 大学院・獣医学研究科, 助教授 (20176895) 松田彰北海道大学, 大学院・薬学研究科, 教授 (90157313) 稲波修北海道大学, 大学院・獣医学研究科, 助教授 (10193559) 岡田幸助岩手大学, 農学部, 教授 (50002077) 佐藤れえ子岩手大学, 農学部, 助教授 (80142892)
Keywords	白血病 / リンパ腫 / 制癌剤 / アポトーシス / カスパーゼ / 細胞死 / ネクローシス / 株化ガン細胞
Research Abstract	本研究は、アポトーシスによる細胞死を効率的に引き起こすことを目指したヌクレオシド系増感剤を開発し、獣医領域で問題となるネコ白血病、イヌリンパ腫などの治療のための制がん剤の開発を目的とする。本年度は、計画に用いるモデル細胞としてヒト胃癌細胞MKN45細胞、チャイニーズハムスターV79細胞ならびに白血病由来細胞MOLT-4細胞の制がん剤による細胞死の機構を検討した。制がん剤としては1-(3-C-ethynyl-beta-D-ribo-pentofuranosyl)cytosine(Ecyd)と2'-chrolo-deoxyadenosine(Cl-dAde)を化学合成し、それぞれ検討した。その結果、両細胞ともそれぞれの制がん剤でカスパーゼファミリーの活性化を伴うアポトーシスを効率よく引き起こすことが明らかとなった。そのメカニズムには細胞によって差があり、MKN45細胞についてはp53依存性のアポトーシスであり、MOLT-4の場合はp53非依存性、Fas活性化依存性のアポトーシスであることが明らかとなった。また、MKN-45細胞の場合は蛋白合成非依存性でカスパーゼ3依存性アポトーシスであること、MOLT-4細胞の場合は、アポトーシス誘導に蛋白質合成を必要とし、FASの活性化、カスパーゼ8を活性化し、カスパーゼ3を活性化しアポトーシスを引き起こすことも明らかとなった。これら結果は、細胞種に依存してアポトーシスを引き起こすシグナル経路の違いがあることを意味している。また、放射線照射では主ににネクローシスを引き起こすが、細胞死を起こさない少量のEcydで処理した細胞に放射線照射するとネクローシスを起こさず、効率よくアポトーシスを誘導させうることもMKN45細胞とV79細胞で明らかとなった。動物細胞でも、イヌのリンパ腫由来癌細胞でのECydとCl-dAdeについて検討したところ、これらの制癌剤は効率よくアポトーシスを起こし、臨床的な応用しうることが明らかとなった。

Research Products

(19 results)

All Other

All Publications (19 results)

[Publications] I.Kashiwakur ら: "Radiation sensitivity of megakaryocyte colony-forming cells in human placental and umbilical cord blood"Radiation Research. 153. 144-152 (2000)
[Publications] Y.Iida ら: "Unique response of double-stranded oligonucleotides containing a single-8-oxo-7,8-dihydroguanine to γ-rays in the frozen aqueous state at 77K"Radiation Research. 154. 582-589 (2000)
[Publications] M.Tsuji ら: "Neuroprotective effect of α-phenyl-N-tert-butylnitrone in gerbil hippocampus is mediated by the mitogen activated protein kinase pathway and heat shock proteins"Neuroscience Letters. 282. 41-44 (2000)
[Publications] Y.Nomura ら: "2-Chloro-2'-deoxyadenosine induces apoptosis through the Fas/Fas ligand pathway in human leukemia cell line MOLT-4"Leukemia. 14. 299-306 (2000)
[Publications] Y.Cui ら: "FMLP-induced formation of F-actin in HL60 cells is dependent on PI3-K but not on intracellular Ca^<2+>, PKC, ERK or p38MAPK"Inflammation Research. 49. 684-691 (2000)
[Publications] O.Inanami ら: "ESR detection of intraphagosomal superoxide in polymorphonuclear leukocytes using 5-(diethoxyphosphoryl)-5-methyl-l-pyrroline-N-oxide"Free Radical Research. 34. 81-92 (2000)
[Publications] M.Yamagutchi ら: "Serum-free coculture system for ex vivo expansion of human cord blood primitive progenitors and SCID mouse-reconstituting cells using human bone marrow primary stromal cells"Experimental Hematology. 29. 174-182 (2001)
[Publications] T.Yamamori ら: "Roles of p38 MAPK, PKC and PI3-K in the signaling pathways of NADPH oxidase activation and phagocytosis in bovine polymorphonuclear leukocytes"FEBS Letter. 467. 253-258 (2001)
[Publications] M.Tsuji ら: "Induction of neurite outgrowth in PC12 cells by α-phenyl-N-tert-butylnitron through activation of protein kinase C and the Ras-extracellular signal-regulated kinase pathway"Journal of Biological Chemistry. 276. 9400-9605 (2001)
[Publications] K.Iwasaki ら: "Kuwabara and M.Niiyama, Isolation, characterization, and cDNA cloning of chicken turpentine-induced protein, a new member of the scavenger receptor cysteine-rich (SRCR) family of protein"Journal of Biological Chemistry. 276. 9400-9405 (2001)
[Publications] H.Abe ら: "Superoxide generation from human polymorphonuclear leukocytes by liposome encapsulated hemoglobin"Artif Cells Blood Substit Immobil Biotechnol. 29. 275-283 (2001)
[Publications] K.Niwa ら: "p38 MAPK and Ca^<2+> contribute to hydrogen peroxide-induced increase of permeability in vascular endotherial cells but ERK does not"Free Radical Research. 35. 519-527 (2001)
[Publications] T.Asanuma ら: "Three dimentional magnetic resonance imaging of lung and liver tumors in mice by use of transversal multislice magnetic resonance image"Comparative Medicine. 51. 138-144 (2001)
[Publications] I.Kashiwakura ら: "Effects of amifostine on the proliferation and differentiation of megakaryocytic progenitor cells"Europearn Journal of Pharmacology. 437. 19-25 (2002)
[Publications] K.Takahashi ら: "Protein synthesis-dependent apoptotic signalling pathway in X-irradiated MOLT-4 human leukaemia cell.line"International Journal of Radiation Biology. 78. 115-124 (2002)
[Publications] O.Inanami ら: "Hypoxia and etanidazol alter radiation-induced apoptosis in HL60 cells but not MOLT-4 cells"International Journal of Radiation Biology. (印刷中).
[Publications] 稲浪修ら: "アポトーシス伝達経路におけるフリーラジカル"細胞. 32. 11-15 (2000)
[Publications] 稲浪修ら: "酸化ストレスと細胞内シグナル伝達"放射線科学. 43. 104-112 (2000)
[Publications] 桑原幹典: "一重項酸素により誘導されるアポトーシスシグナル伝達"医学のあゆみ. 別冊. 86-88 (2001)

2001 Fiscal Year Annual Research Report

アポトーシス情報伝達に着目した新規ヌクレオシド系制癌剤の開発-ネコ白血病、イヌリンパ腫への応用-

Principal Investigator

桑原 幹典 北海道大学, 大学院・獣医学研究科, 教授 (10002081)

Research Products

[Publications] I.Kashiwakur ら: "Radiation sensitivity of megakaryocyte colony-forming cells in human placental and umbilical cord blood"Radiation Research. 153. 144-152 (2000)

[Publications] Y.Iida ら: "Unique response of double-stranded oligonucleotides containing a single-8-oxo-7,8-dihydroguanine to γ-rays in the frozen aqueous state at 77K"Radiation Research. 154. 582-589 (2000)

[Publications] M.Tsuji ら: "Neuroprotective effect of α-phenyl-N-tert-butylnitrone in gerbil hippocampus is mediated by the mitogen activated protein kinase pathway and heat shock proteins"Neuroscience Letters. 282. 41-44 (2000)

[Publications] Y.Nomura ら: "2-Chloro-2'-deoxyadenosine induces apoptosis through the Fas/Fas ligand pathway in human leukemia cell line MOLT-4"Leukemia. 14. 299-306 (2000)

[Publications] Y.Cui ら: "FMLP-induced formation of F-actin in HL60 cells is dependent on PI3-K but not on intracellular Ca^<2+>, PKC, ERK or p38MAPK"Inflammation Research. 49. 684-691 (2000)

[Publications] O.Inanami ら: "ESR detection of intraphagosomal superoxide in polymorphonuclear leukocytes using 5-(diethoxyphosphoryl)-5-methyl-l-pyrroline-N-oxide"Free Radical Research. 34. 81-92 (2000)

[Publications] M.Yamagutchi ら: "Serum-free coculture system for ex vivo expansion of human cord blood primitive progenitors and SCID mouse-reconstituting cells using human bone marrow primary stromal cells"Experimental Hematology. 29. 174-182 (2001)

[Publications] T.Yamamori ら: "Roles of p38 MAPK, PKC and PI3-K in the signaling pathways of NADPH oxidase activation and phagocytosis in bovine polymorphonuclear leukocytes"FEBS Letter. 467. 253-258 (2001)

[Publications] M.Tsuji ら: "Induction of neurite outgrowth in PC12 cells by α-phenyl-N-tert-butylnitron through activation of protein kinase C and the Ras-extracellular signal-regulated kinase pathway"Journal of Biological Chemistry. 276. 9400-9605 (2001)

[Publications] K.Iwasaki ら: "Kuwabara and M.Niiyama, Isolation, characterization, and cDNA cloning of chicken turpentine-induced protein, a new member of the scavenger receptor cysteine-rich (SRCR) family of protein"Journal of Biological Chemistry. 276. 9400-9405 (2001)

[Publications] H.Abe ら: "Superoxide generation from human polymorphonuclear leukocytes by liposome encapsulated hemoglobin"Artif Cells Blood Substit Immobil Biotechnol. 29. 275-283 (2001)

[Publications] K.Niwa ら: "p38 MAPK and Ca^<2+> contribute to hydrogen peroxide-induced increase of permeability in vascular endotherial cells but ERK does not"Free Radical Research. 35. 519-527 (2001)

[Publications] T.Asanuma ら: "Three dimentional magnetic resonance imaging of lung and liver tumors in mice by use of transversal multislice magnetic resonance image"Comparative Medicine. 51. 138-144 (2001)

[Publications] I.Kashiwakura ら: "Effects of amifostine on the proliferation and differentiation of megakaryocytic progenitor cells"Europearn Journal of Pharmacology. 437. 19-25 (2002)

[Publications] K.Takahashi ら: "Protein synthesis-dependent apoptotic signalling pathway in X-irradiated MOLT-4 human leukaemia cell.line"International Journal of Radiation Biology. 78. 115-124 (2002)

[Publications] O.Inanami ら: "Hypoxia and etanidazol alter radiation-induced apoptosis in HL60 cells but not MOLT-4 cells"International Journal of Radiation Biology. (印刷中).

[Publications] 稲浪 修 ら: "アポトーシス伝達経路におけるフリーラジカル"細胞. 32. 11-15 (2000)

[Publications] 稲浪 修 ら: "酸化ストレスと細胞内シグナル伝達"放射線科学. 43. 104-112 (2000)

[Publications] 桑原幹典: "一重項酸素により誘導されるアポトーシスシグナル伝達"医学のあゆみ. 別冊. 86-88 (2001)

桑原幹典北海道大学, 大学院・獣医学研究科, 教授 (10002081)

[Publications] 稲浪修ら: "アポトーシス伝達経路におけるフリーラジカル"細胞. 32. 11-15 (2000)

[Publications] 稲浪修ら: "酸化ストレスと細胞内シグナル伝達"放射線科学. 43. 104-112 (2000)