1990 年度実績報告書

発がんプロモ-ションの機構とその阻害

研究課題

研究課題/領域番号	02151077
研究機関	国立がんセンター
研究代表者	藤木博太国立がんセンター研究所, がん予防研究部, 部長 (60124426)
研究分担者	深見泰夫神戸大学, 遺伝子実験施設, 助教授 (00156746) 橋本祐一東京大学, 応用微生物研究所, 助教授 (90164798) 野瀬清東京大学, 医科学研究所・癌細胞, 助教授 (70012747) 大内和雄東北大学, 薬学部・病態生化学, 教授 (20006357) 石川隆俊東京大学, 医学部・病理, 教授 (30085633)
キーワード	オカダ酸 / オカダ酸クラス発がんプロモ-タ- / ミクロチスチン / 蛋白質脱リン酸化酵素2A / プロスタグランジンE_2 / protein kinase C / CNーTPBP
研究概要	新しい発がんプロモ-タ-、オカダ酸及びオカダ酸クラスの化合物を用いて、プロモ-ションの機構を解明している。オカダ酸クラスの発がんプロモ-タ-は、いづれも、そのレセプタ-である蛋白質脱リン酸化酵素1と2Aに結合し、その活性を抑制した。今回、光親和性オカダ酸誘導体, ^3Hーmethyl7ー0ー(4ーazidobenzoyl)okadaateを用い、精製した蛋白質脱リン酸化酵素2Aと反応させた。この精製した酵素は65kと42kの調節サブユニットと37kの触媒サブユニットからできている。反応液をSDSーPAGEで調べると、 ^3Hーアダクトは触媒サブユニットにのみ認められた。この結果から、オカダ酸は触媒サブユニットに結合し、その活性を抑制することが明らかになった。オカダ酸はNーmethylーN'ーnitroーNーnitroーsoguanidineでイニシエ-トしたラット腺胃に対し、微量で発がんプロモ-ション活性を示した。ミクロチスチンはdiethylnitrosamineでイニシエ-トしたラット肝に対し、微量で発がんプロモ-ション活性を示した。従って、脱リン酸化酵素の阻害を介するオカダ酸クラスの発がんプロモ-ション機構は、いろいろな臓器に共通することが証明された。NIHー3T3細胞と活性化したcーHー<ras>___ーをトランスフェクトしたF25ー15ー5細胞について、発がんプロモ-タ-によるプロスタグランジンE_2産生を比較検討すると,F25ー15ー5細胞の方が強く亢進していた。酵母に含まれる新しいタイプのprotein kinase Cを分離した。この酵素はジアシルグリセロ-ルの代わりに、ホルボ-ルエステルを加えても、活性化が認められなかった。TPAータイププロモ-タ-に特異的に結合する蛋白質(CNーTPBP)は細胞質でheat shock蛋白質90kDと複合体を形成する70kDの蛋白質であり、TPA処理により速やかに核へ移行することが見いだされた。3種のレチノイドレセプタ-は必ずしもmRNAレベルの分布と一致せず、それぞれ固有の役割を担うことが示唆された。

研究成果

(21件)

すべてその他

すべて文献書誌 (21件)

[文献書誌] Suganuma,M.,Fujiki,H.,et al.: "Calyculin A,an inhibitor of protein phophatases,a potent tumor promoter on CDー1 mouse skin." Cancer Res.50. 3521-3525 (1990)
[文献書誌] Nam,KーY.,Fujiki,H.,et al.: "Permeability of a nonーTPAーtype tumor promoter,okadaic acid,through lipid bilayer membrane." Carcinogenesis. 11. 1171-1174 (1990)
[文献書誌] Yoshizawa,S.,Fujiki,H.,et al.: "Tumorーpromoting activity of staurosporine,a protein kinese inhibitor on mouse skin." Cancer Res.50. 4974-4978 (1990)
[文献書誌] Nishiwaki,S.,Fujiki,H.,et al.: "Photoaffinity labeling of protein phosphatase 2A,the receptor for a tumor promoter okadaic acid,by(27ー^3H)methyl 7ーOー(4ーazidobenzoyl)okadaate." Biochem.Biophys.Res.Commun.170. 1359-1364 (1990)
[文献書誌] Nishiwaki,S.,Fujiki,H.,et al.: "Stuructureーactivity relationship within a series of okadaic acid derivatives." Carcinogenesis. 11. 1837-1841 (1990)
[文献書誌] Yoshizawa,S.,Fujiki,H.,et al.: "Inhibition of protein phophatases by microcystin and nodularin associated with hepatotoxicity." J.Cancer Res.Clin.Oncol.116. 609-614 (1990)
[文献書誌] Hashimoto,Y.,et al.: "Cytosolicーnuclear tumor promoterーspecific binding protein (CNーTPBP) in human promyelocystic leukemia cells HLー60." Biochem.Biophys.Res.Commun.166. 1126-1132 (1990)
[文献書誌] Hashimoto,Y.,et al.: "Expression of retinoic acid receptor genes and the ligandーbinding selectivity of retinoic acid receptors(RAR's)." Biochem.Biophys.Res.Commun.116. 1300-1307 (1990)
[文献書誌] Sato,K.,Fukami,Y.,et al.: "A synthetic peptide corresponding to residues 137 to 157 of p60 Vーsrc inhibits tyrosineーspecific protein kinases." Biochem.Biophys.Res.Commun.171. 1152-1159 (1990)
[文献書誌] Saitoh,A.,Izawa,M.,et al.: "Most tumors in transgenic mice with human cーHaー<ras>___ー gene contained somatically activated transgenes." Oncogene. 5. 1195-1200 (1990)
[文献書誌] Hashida,T.,Yasumoto,S.: "Casein kinase II activities related to hyperーphosphoryーlation of human papillomavirus type 16ーE7 oncoーprotein in epidermal keratinocytes." Biochem.Biophys.Res.Commun.172. 958-964 (1990)
[文献書誌] Nakadate,T.,Nishikawa,k.,et al.: "Hー7,a protein kinase C inhibitor,inhibits phorbol esterーcaused ornithine decarboxylase induction but fails to inhibit phorbol esterーcaused suppression of epidermal growth factor binding in primary cultured mouse epidermal cells." Mol.Pharmacol.36. 917-924 (1989)
[文献書誌] Nemoto,N.,Ishikawa,T.,et al.: "Acquisition of aryl hydrocarbon hydroxylase inducibility by aromatic hydrocarbons in monolayerーcyltured hepatocytes from nonresponsive mouse strain." Cancer Res.50. 3226-3236 (1990)
[文献書誌] Nakatsuru,Y.,Ishikawa,T.,et al.: "Immunological detection of <in>___ー <vivo>___ー aflatoxin B_1ーDNA adduct formation in rats,rainbow trout and coho salmon." Carcinogenesis. 11. 1523-1526 (1990)
[文献書誌] Watanabe,M.,Fujiki,H.,Ohuchi,K.,et al.: "Dual effects of staurosporine on arachidonic acid metabolism in rat peritoneal macrophages." Biochem.Biophys.Acta. 1047. 141-147 (1990)
[文献書誌] Nose,K.,et al.: "Abolishment of <cーfos>___ー inducibility in <ras>___ーーtranformed mouse osteoblastic cell lines." Mol.Carcinogenesis. 2. 208-216 (1990)
[文献書誌] Nose,K.,et al.: "Isolation of a gene sequence induced later by tumor promoting 12ーOーtetradecanoylーphorbolー13ーacetate in mouse obteoblastic cells (MC3T3) and expressed constitutively in <ras>___ーーtransformed cells." Cell Growth & Defferentiation. 1. 511-518 (1990)
[文献書誌] Hashimoto,Y.,et al.: "The retionic acid receptors α and β are expressed in the human promyelocytic leukemia cell line HLー60." Molecular Endocrinology. 3. 1046-1052 (1989)
[文献書誌] Fujiki,H.,et al.: "New Tumor promoters from marine sources:The okadaic acid class.in “Mycotoxins and Phycotoxins '88"" Elsevier,Holland, 8 (1989)
[文献書誌] Fujiki,H.,et al.: "A new pathway of tumor promotion by the okadaic acid class compounds.in “The Biology and Medicine of Singnal Transduction"" Raven Press, 5 (1990)
[文献書誌] H.Fujiki,et al.: "A New Pathway of Tumor Promotion by the Okadaic acid class Compounds in Advances in Second Messenger and phosphoprotein Research" Y.Nishizuka,M.Endo,and C.Tanaka, 340-344 (1990)

1990 年度実績報告書

発がんプロモ-ションの機構とその阻害

研究代表者

藤木博太国立がんセンター研究所, がん予防研究部, 部長 (60124426)

研究成果

[文献書誌] Suganuma,M.,Fujiki,H.,et al.: "Calyculin A,an inhibitor of protein phophatases,a potent tumor promoter on CDー1 mouse skin." Cancer Res.50. 3521-3525 (1990)

[文献書誌] Nam,KーY.,Fujiki,H.,et al.: "Permeability of a nonーTPAーtype tumor promoter,okadaic acid,through lipid bilayer membrane." Carcinogenesis. 11. 1171-1174 (1990)

[文献書誌] Yoshizawa,S.,Fujiki,H.,et al.: "Tumorーpromoting activity of staurosporine,a protein kinese inhibitor on mouse skin." Cancer Res.50. 4974-4978 (1990)

[文献書誌] Nishiwaki,S.,Fujiki,H.,et al.: "Photoaffinity labeling of protein phosphatase 2A,the receptor for a tumor promoter okadaic acid,by(27ー^3H)methyl 7ーOー(4ーazidobenzoyl)okadaate." Biochem.Biophys.Res.Commun.170. 1359-1364 (1990)

[文献書誌] Nishiwaki,S.,Fujiki,H.,et al.: "Stuructureーactivity relationship within a series of okadaic acid derivatives." Carcinogenesis. 11. 1837-1841 (1990)

[文献書誌] Yoshizawa,S.,Fujiki,H.,et al.: "Inhibition of protein phophatases by microcystin and nodularin associated with hepatotoxicity." J.Cancer Res.Clin.Oncol.116. 609-614 (1990)

[文献書誌] Hashimoto,Y.,et al.: "Cytosolicーnuclear tumor promoterーspecific binding protein (CNーTPBP) in human promyelocystic leukemia cells HLー60." Biochem.Biophys.Res.Commun.166. 1126-1132 (1990)

[文献書誌] Hashimoto,Y.,et al.: "Expression of retinoic acid receptor genes and the ligandーbinding selectivity of retinoic acid receptors(RAR's)." Biochem.Biophys.Res.Commun.116. 1300-1307 (1990)

[文献書誌] Sato,K.,Fukami,Y.,et al.: "A synthetic peptide corresponding to residues 137 to 157 of p60 Vーsrc inhibits tyrosineーspecific protein kinases." Biochem.Biophys.Res.Commun.171. 1152-1159 (1990)

[文献書誌] Saitoh,A.,Izawa,M.,et al.: "Most tumors in transgenic mice with human cーHaー<ras>___ー gene contained somatically activated transgenes." Oncogene. 5. 1195-1200 (1990)

[文献書誌] Hashida,T.,Yasumoto,S.: "Casein kinase II activities related to hyperーphosphoryーlation of human papillomavirus type 16ーE7 oncoーprotein in epidermal keratinocytes." Biochem.Biophys.Res.Commun.172. 958-964 (1990)

[文献書誌] Nemoto,N.,Ishikawa,T.,et al.: "Acquisition of aryl hydrocarbon hydroxylase inducibility by aromatic hydrocarbons in monolayerーcyltured hepatocytes from nonresponsive mouse strain." Cancer Res.50. 3226-3236 (1990)

[文献書誌] Nakatsuru,Y.,Ishikawa,T.,et al.: "Immunological detection of <in>_ー <vivo>_ー aflatoxin B_1ーDNA adduct formation in rats,rainbow trout and coho salmon." Carcinogenesis. 11. 1523-1526 (1990)

[文献書誌] Watanabe,M.,Fujiki,H.,Ohuchi,K.,et al.: "Dual effects of staurosporine on arachidonic acid metabolism in rat peritoneal macrophages." Biochem.Biophys.Acta. 1047. 141-147 (1990)

[文献書誌] Nose,K.,et al.: "Abolishment of <cーfos>_ー inducibility in <ras>_ーーtranformed mouse osteoblastic cell lines." Mol.Carcinogenesis. 2. 208-216 (1990)

[文献書誌] Nose,K.,et al.: "Isolation of a gene sequence induced later by tumor promoting 12ーOーtetradecanoylーphorbolー13ーacetate in mouse obteoblastic cells (MC3T3) and expressed constitutively in <ras>___ーーtransformed cells." Cell Growth & Defferentiation. 1. 511-518 (1990)

[文献書誌] Hashimoto,Y.,et al.: "The retionic acid receptors α and β are expressed in the human promyelocytic leukemia cell line HLー60." Molecular Endocrinology. 3. 1046-1052 (1989)

[文献書誌] Fujiki,H.,et al.: "New Tumor promoters from marine sources:The okadaic acid class.in “Mycotoxins and Phycotoxins '88"" Elsevier,Holland, 8 (1989)

[文献書誌] Fujiki,H.,et al.: "A new pathway of tumor promotion by the okadaic acid class compounds.in “The Biology and Medicine of Singnal Transduction"" Raven Press, 5 (1990)

[文献書誌] H.Fujiki,et al.: "A New Pathway of Tumor Promotion by the Okadaic acid class Compounds in Advances in Second Messenger and phosphoprotein Research" Y.Nishizuka,M.Endo,and C.Tanaka, 340-344 (1990)

1990 年度 実績報告書

発がんプロモ-ションの機構とその阻害

研究代表者

藤木 博太 国立がんセンター研究所, がん予防研究部, 部長 (60124426)

研究成果

[文献書誌] Suganuma,M.,Fujiki,H.,et al.: "Calyculin A,an inhibitor of protein phophatases,a potent tumor promoter on CDー1 mouse skin." Cancer Res.50. 3521-3525 (1990)

[文献書誌] Nam,KーY.,Fujiki,H.,et al.: "Permeability of a nonーTPAーtype tumor promoter,okadaic acid,through lipid bilayer membrane." Carcinogenesis. 11. 1171-1174 (1990)

[文献書誌] Yoshizawa,S.,Fujiki,H.,et al.: "Tumorーpromoting activity of staurosporine,a protein kinese inhibitor on mouse skin." Cancer Res.50. 4974-4978 (1990)

[文献書誌] Nishiwaki,S.,Fujiki,H.,et al.: "Photoaffinity labeling of protein phosphatase 2A,the receptor for a tumor promoter okadaic acid,by(27ー^3H)methyl 7ーOー(4ーazidobenzoyl)okadaate." Biochem.Biophys.Res.Commun.170. 1359-1364 (1990)

[文献書誌] Nishiwaki,S.,Fujiki,H.,et al.: "Stuructureーactivity relationship within a series of okadaic acid derivatives." Carcinogenesis. 11. 1837-1841 (1990)

[文献書誌] Yoshizawa,S.,Fujiki,H.,et al.: "Inhibition of protein phophatases by microcystin and nodularin associated with hepatotoxicity." J.Cancer Res.Clin.Oncol.116. 609-614 (1990)

[文献書誌] Hashimoto,Y.,et al.: "Cytosolicーnuclear tumor promoterーspecific binding protein (CNーTPBP) in human promyelocystic leukemia cells HLー60." Biochem.Biophys.Res.Commun.166. 1126-1132 (1990)

[文献書誌] Hashimoto,Y.,et al.: "Expression of retinoic acid receptor genes and the ligandーbinding selectivity of retinoic acid receptors(RAR's)." Biochem.Biophys.Res.Commun.116. 1300-1307 (1990)

[文献書誌] Sato,K.,Fukami,Y.,et al.: "A synthetic peptide corresponding to residues 137 to 157 of p60 Vーsrc inhibits tyrosineーspecific protein kinases." Biochem.Biophys.Res.Commun.171. 1152-1159 (1990)

[文献書誌] Saitoh,A.,Izawa,M.,et al.: "Most tumors in transgenic mice with human cーHaー<ras>___ー gene contained somatically activated transgenes." Oncogene. 5. 1195-1200 (1990)

[文献書誌] Hashida,T.,Yasumoto,S.: "Casein kinase II activities related to hyperーphosphoryーlation of human papillomavirus type 16ーE7 oncoーprotein in epidermal keratinocytes." Biochem.Biophys.Res.Commun.172. 958-964 (1990)

[文献書誌] Nemoto,N.,Ishikawa,T.,et al.: "Acquisition of aryl hydrocarbon hydroxylase inducibility by aromatic hydrocarbons in monolayerーcyltured hepatocytes from nonresponsive mouse strain." Cancer Res.50. 3226-3236 (1990)

[文献書誌] Nakatsuru,Y.,Ishikawa,T.,et al.: "Immunological detection of <in>___ー <vivo>___ー aflatoxin B_1ーDNA adduct formation in rats,rainbow trout and coho salmon." Carcinogenesis. 11. 1523-1526 (1990)

[文献書誌] Watanabe,M.,Fujiki,H.,Ohuchi,K.,et al.: "Dual effects of staurosporine on arachidonic acid metabolism in rat peritoneal macrophages." Biochem.Biophys.Acta. 1047. 141-147 (1990)

[文献書誌] Nose,K.,et al.: "Abolishment of <cーfos>___ー inducibility in <ras>___ーーtranformed mouse osteoblastic cell lines." Mol.Carcinogenesis. 2. 208-216 (1990)

[文献書誌] Nose,K.,et al.: "Isolation of a gene sequence induced later by tumor promoting 12ーOーtetradecanoylーphorbolー13ーacetate in mouse obteoblastic cells (MC3T3) and expressed constitutively in <ras>___ーーtransformed cells." Cell Growth & Defferentiation. 1. 511-518 (1990)

[文献書誌] Hashimoto,Y.,et al.: "The retionic acid receptors α and β are expressed in the human promyelocytic leukemia cell line HLー60." Molecular Endocrinology. 3. 1046-1052 (1989)

[文献書誌] Fujiki,H.,et al.: "New Tumor promoters from marine sources:The okadaic acid class.in “Mycotoxins and Phycotoxins '88"" Elsevier,Holland, 8 (1989)

[文献書誌] Fujiki,H.,et al.: "A new pathway of tumor promotion by the okadaic acid class compounds.in “The Biology and Medicine of Singnal Transduction"" Raven Press, 5 (1990)

[文献書誌] H.Fujiki,et al.: "A New Pathway of Tumor Promotion by the Okadaic acid class Compounds in Advances in Second Messenger and phosphoprotein Research" Y.Nishizuka,M.Endo,and C.Tanaka, 340-344 (1990)

1990 年度実績報告書

藤木博太国立がんセンター研究所, がん予防研究部, 部長 (60124426)

[文献書誌] Nakatsuru,Y.,Ishikawa,T.,et al.: "Immunological detection of <in>_ー <vivo>_ー aflatoxin B_1ーDNA adduct formation in rats,rainbow trout and coho salmon." Carcinogenesis. 11. 1523-1526 (1990)

[文献書誌] Nose,K.,et al.: "Abolishment of <cーfos>_ー inducibility in <ras>_ーーtranformed mouse osteoblastic cell lines." Mol.Carcinogenesis. 2. 208-216 (1990)