2002 Fiscal Year Annual Research Report

銅トランスポータATP7A・ATP7Bによる抗癌剤耐性の分子機構の解析

Research Project

Project/Area Number	14030072
Research Institution	Kagoshima University
Principal Investigator	古川龍彦鹿児島大学, 医学部, 助教授 (40219100)
Keywords	銅トランスポータ / 抗癌剤耐性 / シスプラチン / PタイプATPase
Research Abstract	複数のATP7A発現細胞を用いて、種々の抗癌剤に対する耐性度をMTTアッセイで調べた。アドリアマイシン(ADM)、Mitxantrone、CPT-11、VP-16、Paclitaxel、CDDP、SN-38に対してそれぞれ10、5.5、3.5、10、2.5、100倍の耐性を示すことが確認できた。抗癌剤ADMに対してATP7A発現細胞は約10倍耐性となる。その蛍光を利用して、ADMのATP7A発現細胞での細胞内の局在を調べた。その結果、親株細胞やATP7B発現細胞では核に強い蛍光が観察されたが、ATP7A発現細胞ではADMの蛍光が全体に弱く、蛍光も核周辺に局在していることが確認された。さらに、RI標識ADMを用いてその細胞外への排出を調べたATP7A発現細胞でADMの排出が亢進して蓄積が減少していた。ヒンジドメインの1270番目のアスパラギンをセリンに置換した変異型ATP7B(激症型ウイルソン病の患者に見られる)の発現細胞と6つある銅結合部位の5種類の欠失変異ATP7B発現細胞を単離して、CDDPに対する耐性を調べた。銅の輸送に重要であると考えられるヒンジドメインの変異体や6番目の銅結合部位の欠失変異体ではCDDPに対して感受性を示し、ATP7Bの輸送機能がCDDP耐性に関わっている可能性をしめした。また、野生型のATP7B発現細胞について様々な発現量を示す10個以上におよぶATP7B発現細胞株と単離してCDDP耐性を検討した。その結果ATP7Bの発現量が非常に高い場株ではCDDPにむしろ感受性になる結果を得た。 ATP7AとATP7B発現細胞の耐性克服薬剤のスクリーニングを行っているが、現在までのところ、有望な薬剤は見いだしていない。

Research Products
(12 results)

All Other

All Publications (12 results)

[Publications] Ren XQ.: "A Positively Charged Amino Acid proximate to C-terminus of TM17 of MRP1 is indispensable for GSH-dependent binding of substrates and for transport of LTC4"Biochemistry. 41. 14132-14140 (2002)
[Publications] Ren XQ.: "Localization of the GSH-dependent photolabling site of an agosterol A analog on human MRP1"British Journal of Pharmacology. (印刷中). (2003)
[Publications] Wang X: "Breast cancer resistance protein (BCRP/ABCG2) induces cellular resistance to HIV-1 nucleoside reverse transcriptase inhibitors"Mol.Pharmacology. 63. 65-72 (2003)
[Publications] Che X-F.: "Reversal of P-glycoprotein mediated multidrug resistance by newly synthesized 1,4-bensothiazipine derivative, JTV-519"Cancer Lett.. 187. 111-119 (2002)
[Publications] Haraguchi M.: "Targeted Deletion of both Thymidine Phosphorylase and Uridine Phosphorylase and consequent disorders in mice"Mol.Cell.Biol.. 22. 5212-5221 (2002)
[Publications] Uchimiya H.: "Suppression of thymidin e phosphorylase-mediated angiogenesis and tumor growth by 2-deoxy-L-ribose"Cancer Res.. 62. 2834-2939 (2002)
[Publications] Tsujikawa K.: "Regulation of Lck and Fyn Tyrosine Kinase Activities by Transmembrane Protein Tyrosine Phosphatase Leukocyte Common Antigen-Related Molecule"Mol.Cancer Res.. 1. 155-163 (2002)
[Publications] Ikeda R.: "Molecular basis for the inhibition of hypoxia-induced apoptosis by 2-deoxy-D-ribose"Biochem.Biophys.Res.Commun.. 291. 806-812 (2003)
[Publications] Ikeda R.: "Thymidine phosphorylase inhibits apoptosis induced cisplatin"Biochem.Biophys.Res.Commun.. 301. 358-363 (2003)
[Publications] Aoki S.: "Lembehsterols A and B, novel sulfated sterols inhibiting thyinidine phosphorylase, from the marine sponge Petrosia strongylata"Chem.Pharm.Bull.. 50. 827-830 (2002)
[Publications] Mukai M.: "Enhanced nucleiotide excision repair in cisplatin resistant human KB carcinoma cells"Oncology Report. 9. 839-844 (2002)
[Publications] Ohno N.: "Augmentation of the cytotoxic activity of an immunotoxin, anti-Tac(Fv)-PE40KDEL, in adutl T leukemia cells"Leukemia Lymphoma. 43. 885-888 (2002)

2002 Fiscal Year Annual Research Report

銅トランスポータATP7A・ATP7Bによる抗癌剤耐性の分子機構の解析

Principal Investigator

古川 龍彦 鹿児島大学, 医学部, 助教授 (40219100)

Research Products

[Publications] Ren XQ.: "A Positively Charged Amino Acid proximate to C-terminus of TM17 of MRP1 is indispensable for GSH-dependent binding of substrates and for transport of LTC4"Biochemistry. 41. 14132-14140 (2002)

[Publications] Ren XQ.: "Localization of the GSH-dependent photolabling site of an agosterol A analog on human MRP1"British Journal of Pharmacology. (印刷中). (2003)

[Publications] Wang X: "Breast cancer resistance protein (BCRP/ABCG2) induces cellular resistance to HIV-1 nucleoside reverse transcriptase inhibitors"Mol.Pharmacology. 63. 65-72 (2003)

[Publications] Che X-F.: "Reversal of P-glycoprotein mediated multidrug resistance by newly synthesized 1,4-bensothiazipine derivative, JTV-519"Cancer Lett.. 187. 111-119 (2002)

[Publications] Haraguchi M.: "Targeted Deletion of both Thymidine Phosphorylase and Uridine Phosphorylase and consequent disorders in mice"Mol.Cell.Biol.. 22. 5212-5221 (2002)

[Publications] Uchimiya H.: "Suppression of thymidin e phosphorylase-mediated angiogenesis and tumor growth by 2-deoxy-L-ribose"Cancer Res.. 62. 2834-2939 (2002)

[Publications] Tsujikawa K.: "Regulation of Lck and Fyn Tyrosine Kinase Activities by Transmembrane Protein Tyrosine Phosphatase Leukocyte Common Antigen-Related Molecule"Mol.Cancer Res.. 1. 155-163 (2002)

[Publications] Ikeda R.: "Molecular basis for the inhibition of hypoxia-induced apoptosis by 2-deoxy-D-ribose"Biochem.Biophys.Res.Commun.. 291. 806-812 (2003)

[Publications] Ikeda R.: "Thymidine phosphorylase inhibits apoptosis induced cisplatin"Biochem.Biophys.Res.Commun.. 301. 358-363 (2003)

[Publications] Aoki S.: "Lembehsterols A and B, novel sulfated sterols inhibiting thyinidine phosphorylase, from the marine sponge Petrosia strongylata"Chem.Pharm.Bull.. 50. 827-830 (2002)

[Publications] Mukai M.: "Enhanced nucleiotide excision repair in cisplatin resistant human KB carcinoma cells"Oncology Report. 9. 839-844 (2002)

[Publications] Ohno N.: "Augmentation of the cytotoxic activity of an immunotoxin, anti-Tac(Fv)-PE40KDEL, in adutl T leukemia cells"Leukemia Lymphoma. 43. 885-888 (2002)

古川龍彦鹿児島大学, 医学部, 助教授 (40219100)