1990 Fiscal Year Annual Research Report

新しい制がん剤耐性遺伝子群の解析と克服法の確立

Research Project

Project/Area Number	02151053
Research Institution	大分医科大学
Principal Investigator	桑野信彦大分医科大学, 医学部, 教授 (80037431)
Co-Investigator(Kenkyū-buntansha)	稲葉実癌研究会, 癌化学療法センター, 主任研究員 (60085636) 河野公俊大分医科大学, 医学部, 助教授 (00153479) 杉本芳一癌研究会, 癌化学療法センター, 研究員 (10179161) 植田和光京都大学, 農学部, 助手 (10151789) 秋山伸一鹿児島大学, 医学部, 教授 (60117413)
Keywords	耐性獲得 / 多剤耐性遺伝子 / Pー糖蛋白 / 耐性克服 / DNAトポイソメラ-ゼ / メタロチオネイン / グルタチオン・チオ-ル・トランスフェラ-ゼ / 5ーフルオロウラシル耐性
Research Abstract	本研究では、制がん剤耐性獲得に関与する機構を分子レベルで解析し腫瘍耐性のマ-カ-としての有用性を検索するとともにその耐性因子を標的にした耐性克服法を考案することを進めた。その結果、1.多剤耐性MDRー1遺伝子及びその産物gp170に関して(1)腎副腎腸などでの発現を制御するエンハンサ-の単離及び副腎のgp170が薬剤結合部位の存在を明らかにした。(2)ヒト血漿中にgp170の生理的輸送蛋白の存在と部分精製を行った。(3)MDRー1遺伝子の発現が砒素処理で上昇し熱ショック蛋白の関与が示唆され、又同プロモ-タ発現は数多くの制がん剤によって上昇する機構を提示した。(4)多剤耐性の獲得時にgp170以外に85kDa蛋白をはじめとした幾つかの蛋白因子及び相当する遺伝子群の単離を進めた。(5)gp170を標的にした克服剤としてジハイドロピリジン系の異なる構造式を示す薬剤の中から培養系ならびに担癌動物実験系で強力な克服作用を示す物質を検出した。(6)ヒト急性白血病細胞を株化し多剤耐性を示す細胞株を分離し、親株耐性株ともヌ-ドマウスへ可移植性の系を樹立した。さらにMDRー1遺伝子が白血病、乳癌、Tー細胞白血病また大腸癌などで上昇している症例がみられ化学療法後に発現頻度が増加した。2.非多剤耐性に関して(1)DNAトポイソメラ-ゼI及びIIの量また質的変化がエトポシドやカンプトテシンに対する感受性を左右させていた。(2)カンプトテシン耐性のトポイソメラ-ゼIに効果を示す薬剤の探索を進めた。(3)5ーフルオロウラシルの殺細胞作用の速度論的解析から5FUR及び5FUdRの両型がその効果発現に関与していることを提示した。ヒト胃癌、結腸癌においてFUR経路が主として機能しており自然耐性の獲得においても大切であることを示唆した。(4)メタロチオネイン遺伝子はシスプラチン耐性卵巣癌で発現が増加しているがアドリアマイシン耐性肺癌では低下していた。(5)シスプラチン耐性にグルタチオンーSートランスフェラ-ゼの上昇が関与した。

Research Products

(21 results)

All Other

All Publications (21 results)

[Publications] Kiue,A.: "Activities of newly synthesized dihydropyridine in overcoming of vincristine resistance,calcium antagonism and inhibition of phoroーaffinity labeling Pーglycoprotein." Cancer Research. 50. 310-317 (1990)
[Publications] Nakagawara,A.: "Inverse correlation between expression of multidrugーresistance gene (mdrー1) and Nーmyc oncogene in human neuroblastoma." Cancer Research. 50. 3043-3047 (1990)
[Publications] Miyazaki,M.: "Drug resistance to cisーdiamminedichloroplatinum(II) in Chinese hamster ovary cell line transfected with glutathione Sーtransferase Pi gene." Biochem.Biophys.Res.Commun.166. 1358-1364 (1990)
[Publications] Mizoguchi,T.: "Expression of the MDR1 gene in human gastric and colorectal carcinomas." J.Nat.Cancer Inst.82. 1679-1683 (1990)
[Publications] Kuwazuru,Y.: "Expression of Pーglycoprotein in adult Tーcell leukemia cells." Blood. 76. 2065-2071 (1990)
[Publications] Shudo,N.: "Two pyridine analogues with more effective ability to reverse multidrug resistance and with lower calcium channel blocking activity than their dihydropyridine counterparts" Cancer Research. 50. 3055-3061 (1990)
[Publications] Chen,C.: "Genomic organization of the human multidrug resistance (MDR1) gene and origin of Pーglycoproteins" J.BIol.Chem.265. 506-514 (1990)
[Publications] Sugawara,I.: "In situ localization of the human multidrugーresistance gene mRNA usingーthymineーthymine dimerized single stranded cDNA." Jpn.J.Cancer Res.81. 949-955 (1990)
[Publications] Tanaka,S.: "Use of recombinant Pーglycoprotein fragments to produce antibodies to the multidrug transfer." Biochem.Biophys.Res.Commun.166. 180-186 (1990)
[Publications] Sugimoto,Y.: "Detection of 300ーkilodalton membrane protein in adriamycinーresistant human tumor cells by a monoclonal antibody MRK18." Biochem.Biophys.Res.Commun.169. 686-691 (1990)
[Publications] Sugimoto,Y.: "Decreased expression of DNA topoisomerase I in camptothecinーresistant tumor cell lines as determined by a monoclonal antibody." Cancer Research. 50. 6925-6930 (1990)
[Publications] Sugimoto,Y.: "Elevated expression of DNA topoisomerase II in camptothecinーresistant human tumor cell lines" Cancer Research. 50. (1990)
[Publications] Kohno,K.: "The direct activation of human multidrug resistance gene (MDR1) by anticancer agents." Biochem.Biophys.Res.Commun.165. 1415-1421 (1989)
[Publications] Matsuo,K.: "Reduction of drug accumulation and DNA topoisomerase II activity in acquired teneposideーresistant human cancer KB cell lines" Cancer Research. 50. 5819-5824 (1990)
[Publications] Kohno,K.: "Tissueーspecific enhancer of the human multidrugーresistance (MDR1) gene" J.Biol.Chem.265. 19690-19696 (1990)
[Publications] Inaba,M.: "Kinetics analysis of 5ーfluorouracil action against various cancer cells" Jpn.J.Cancer Res.81. 1039-1044 (1990)
[Publications] 稲葉実: "Phrmacokineticsからー制がん作用の速度論的解析とその応用" Oncologia. 23. 7-15 (1990)
[Publications] Kunimoto,S.: "Biological activities of new anthracyclines containing fluorine,FAD104 and its merabolites." J.Antibiotics. 43. 556-565 (1990)
[Publications] Ohno,R.: "Effect of granulocyte colony stimulating factor after intensive induction therapy in relapsed or refractory acute leukemia:A randomized controlled study." New Engl.J.Med.323. 871 (1990)
[Publications] Ohno,R.: "An early phase IIーstudy of CPTー11ーa new derivative of camptothecin,for the treatment of leukemia" J.Clinical Oncology. 8. 1907-1912 (1990)
[Publications] 向山雄人: "自家骨髄移植併用超大量化学療法による抗癌剤耐性克服の試み" 癌と化学療法. 16. 2013-2018 (1989)

1990 Fiscal Year Annual Research Report

新しい制がん剤耐性遺伝子群の解析と克服法の確立

Principal Investigator

桑野 信彦 大分医科大学, 医学部, 教授 (80037431)

Research Products

[Publications] Kiue,A.: "Activities of newly synthesized dihydropyridine in overcoming of vincristine resistance,calcium antagonism and inhibition of phoroーaffinity labeling Pーglycoprotein." Cancer Research. 50. 310-317 (1990)

[Publications] Nakagawara,A.: "Inverse correlation between expression of multidrugーresistance gene (mdrー1) and Nーmyc oncogene in human neuroblastoma." Cancer Research. 50. 3043-3047 (1990)

[Publications] Miyazaki,M.: "Drug resistance to cisーdiamminedichloroplatinum(II) in Chinese hamster ovary cell line transfected with glutathione Sーtransferase Pi gene." Biochem.Biophys.Res.Commun.166. 1358-1364 (1990)

[Publications] Mizoguchi,T.: "Expression of the MDR1 gene in human gastric and colorectal carcinomas." J.Nat.Cancer Inst.82. 1679-1683 (1990)

[Publications] Kuwazuru,Y.: "Expression of Pーglycoprotein in adult Tーcell leukemia cells." Blood. 76. 2065-2071 (1990)

[Publications] Shudo,N.: "Two pyridine analogues with more effective ability to reverse multidrug resistance and with lower calcium channel blocking activity than their dihydropyridine counterparts" Cancer Research. 50. 3055-3061 (1990)

[Publications] Chen,C.: "Genomic organization of the human multidrug resistance (MDR1) gene and origin of Pーglycoproteins" J.BIol.Chem.265. 506-514 (1990)

[Publications] Sugawara,I.: "In situ localization of the human multidrugーresistance gene mRNA usingーthymineーthymine dimerized single stranded cDNA." Jpn.J.Cancer Res.81. 949-955 (1990)

[Publications] Tanaka,S.: "Use of recombinant Pーglycoprotein fragments to produce antibodies to the multidrug transfer." Biochem.Biophys.Res.Commun.166. 180-186 (1990)

[Publications] Sugimoto,Y.: "Detection of 300ーkilodalton membrane protein in adriamycinーresistant human tumor cells by a monoclonal antibody MRK18." Biochem.Biophys.Res.Commun.169. 686-691 (1990)

[Publications] Sugimoto,Y.: "Decreased expression of DNA topoisomerase I in camptothecinーresistant tumor cell lines as determined by a monoclonal antibody." Cancer Research. 50. 6925-6930 (1990)

[Publications] Sugimoto,Y.: "Elevated expression of DNA topoisomerase II in camptothecinーresistant human tumor cell lines" Cancer Research. 50. (1990)

[Publications] Kohno,K.: "The direct activation of human multidrug resistance gene (MDR1) by anticancer agents." Biochem.Biophys.Res.Commun.165. 1415-1421 (1989)

[Publications] Matsuo,K.: "Reduction of drug accumulation and DNA topoisomerase II activity in acquired teneposideーresistant human cancer KB cell lines" Cancer Research. 50. 5819-5824 (1990)

[Publications] Kohno,K.: "Tissueーspecific enhancer of the human multidrugーresistance (MDR1) gene" J.Biol.Chem.265. 19690-19696 (1990)

[Publications] Inaba,M.: "Kinetics analysis of 5ーfluorouracil action against various cancer cells" Jpn.J.Cancer Res.81. 1039-1044 (1990)

[Publications] 稲葉 実: "Phrmacokineticsからー制がん作用の速度論的解析とその応用" Oncologia. 23. 7-15 (1990)

[Publications] Kunimoto,S.: "Biological activities of new anthracyclines containing fluorine,FAD104 and its merabolites." J.Antibiotics. 43. 556-565 (1990)

[Publications] Ohno,R.: "Effect of granulocyte colony stimulating factor after intensive induction therapy in relapsed or refractory acute leukemia:A randomized controlled study." New Engl.J.Med.323. 871 (1990)

[Publications] Ohno,R.: "An early phase IIーstudy of CPTー11ーa new derivative of camptothecin,for the treatment of leukemia" J.Clinical Oncology. 8. 1907-1912 (1990)

[Publications] 向山 雄人: "自家骨髄移植併用超大量化学療法による抗癌剤耐性克服の試み" 癌と化学療法. 16. 2013-2018 (1989)

桑野信彦大分医科大学, 医学部, 教授 (80037431)

[Publications] 稲葉実: "Phrmacokineticsからー制がん作用の速度論的解析とその応用" Oncologia. 23. 7-15 (1990)

[Publications] 向山雄人: "自家骨髄移植併用超大量化学療法による抗癌剤耐性克服の試み" 癌と化学療法. 16. 2013-2018 (1989)