• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Development and clinical application of the molecular diagnostic tests for individualization of cancer chemotherapy: detection of gene polymorphism of metabolizing enzyme and expression pattern

Research Project

Project/Area Number 12672254
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Laboratory medicine
Research InstitutionTokai University

Principal Investigator

MIYACHI Hayato  Tokai University, School of Medicine, Associate Professor, 医学部, 助教授 (20174196)

Project Period (FY) 2000 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 2002: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2001: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2000: ¥900,000 (Direct Cost: ¥900,000)
KeywordsLeukemia / Drug resistance / Anti-folate resistance / Molecular diagnostic test / Gene expression / Gene mutation / 癌化学療法 / 耐性 / 遺伝子多型
Research Abstract

Drug resistance is a major obstacle in chemotherapy of leukemia patients. Since the treatment responsiveness is different among patients, the therapy should be tailored to specific subtypes of diseases such as defined by drug resistance. To elucidate mechanism(s) of methotrexate (MTX)-resistance as a possible reason behind treatment failure in high-dose MTX regimens combined with leucovorin (LV) rescue, we established MTX-resistant human T-cell leukemia cell line CCRF-CEM cells in the presence of excessive leucovorin, and characterized their property. Continuous exposure of the cells to escalating concentrations of MTX up to 20μM in the presence of 1000 nM leucovorin resulted in establishment of three MTX-resistant sublines with a wide disparity of resistance degree over 4 logarithmic range by approximately 40-, 900- and 44000-fold, respectively. Transraembrane transport of MTX in these sublines was diminished to 52%, 35% and 12%, respectively. Intracellular retention of MTX in these s … More ublines was not different from the parent cells. A cell growth study in various concentrations of leucovorin showed that cells with higher resistance to MTX required more leucovorin supply for their optimal growth. In parallel with the resistance levels, there was increase in mRNA expression of dihydrofolate reductase gene and decrease in that of thymidylate synthase gene, but no change in that of reduced folate carrier (RFC1) gene, as assessed by northern blot analysis. Sequencing of RFC1 gene in all of the 3 sublines revealed a point mutation in codon 47 (TCC→TTC) resulting in substitution of Phe for Ser residue, and additional deletion of CTG of codon 112 in the subline with the highest resistance. In summary, MTX exposure to CCRF-CEM cells in the presence of 1000 nM leucovorin resulted in an establishment of heterogeneous cell populations with a wide range of transport-mediated MTX-resistance, which was associated with differential alterations of RFC gene. Combination of gene expression and mutations involved in the molecular mechanisms of MTX-resistance may give targets for detection and evaluation of it. Less

Report

(4 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • 2000 Annual Research Report
  • Research Products

    (32 results)

All Other

All Publications (32 results)

  • [Publications] Takemura, Y., Kobayashi, H., Miyachi, H.: "Raltitrexed, a novel folate-based thymidylate synthase inhibitor for the treatment of acute leukemia : is this drug active against acute myelogenous leukemia?"Int J Hematol.. 72. 112-114 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hatanaka H, Abe Y, Naruke M, Asai S, Miyachi H, et al.: "Modulation of multidrug resistance in a cancer cell line by anti-multidrug resistance-associated protein (MRP) ribozyme"Anticancer Res.. 21. 879-886 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Efeerth T, Dunstan H, Sauerbrey A, Miyachi H, et al.: "The anti-malarial artesunate is also active against cancer"Intern J Oncol.. 18. 767-773 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nagata J, Kijima H, Hatanaka H, Asai S, Miyachi H, et al.: "Reversal of cisplatin and multidrug resistance by ribozyme-mediated glutathione suppression"Biochem Biophys Res Com.. 286. 406-413 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kobayashi H, Takemura Y, Miyachi H: "Novel approaches to reversing anti-cancer drug resistance using gene-specific therapeutics"Human Cell. 14. 172-184 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Takemura, Y, Kobayashi, H, Miyachi, H: "Antifolate resistance and its circumvention by new analogues"Human Cell. 14. 185-202 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Efferth T, Miyachi H, Drexler HG, Gebbart E: "Methylthioadenosine phosphorylase as target for chemoselective treatment of acute T-cell lymphoblastic leukemia cells"Blood Cells Mol Dis.. 28. 47-56 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nagata J., Kijima H., Nakahata H., Asai S., Miyachi H., et al.: "Reversal of drug resistance using hammerhead ribozymes against multidrug resistance-associated protein and multidrug resistance 1 gene"Intern J Oncol. 21. 1021-1026 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Miyachi H, et al.: "Gene expression profiling for predicting treatment responsiveness of cancer"Recent Adv Res Updates. 3. 5-12 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 宮地勇人, 他: "cDNAマイクロアレイ技術の臨床検査への応用.遺伝子発現プロファイル解析による癌の薬剤耐性形質の評価"臨床病理. 50. 161-168 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Asai S, Miyachi H, Kobayashi H, Takemura Y, Ando Y.: "A large diversity in transport-mediated methotrexate resisitance in human leuekmia cell line CCRF-CEM established in a high concentration of leucovorin"Cancer Science. (in press). (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Takemura, Y., Kobayashi, H., Miyachi, H.: "Raltitrexed, a novel folate-based thymidylate synthase inhibitor for the treatment of acute leukemia: is this drug active against acute myelogenous leukemia?"Int J Hematol.. 72. 112-114 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hatanaka H, Abe Y, Naruke M, Asai S, Miyachi H, et al.: "Modulation of multidrug resistance in a cancer cell line by anti-multidrug resistance-associated protein (MRP) ribozyme"Anticancer Res.. 21. 879-886 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Efeerth T, Dunstan H, Sauerbrey A, Miyachi H, et al.: "The anti-malarial artesunate is also active against cancer"Intern J Oncol.. 18. 767-773 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nagata J, Kijima H, Hatanaka H, Asai S, Miyachi H, et al.: "Reversal of cisplatin and multidrug resistance by ribozyme-mediated glutathione suppression"Biochem Biophys Res Com. 286. 406-413 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kobayashi H, Takemura Y, Miyachi H: "Novel approaches to reversing anti-cancer drug resistance using gene-specific therapeutics"Human Cell. 14. 172-184 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Takemura Y, Kobayashi H, Miyachi H: "Antifolate resistance and its circumvention by new analogues"Human Cell. 14. 185-2023 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Efferth T, Miyachi H, Drexler HG and Gebhart E: "Methylthioadenosine phosphorylase as target for chemoselective treatment of acute T-cell lymphoblastic leukemia cells"Blood Cells Mol Dis.. 28. 47-5 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nagata J., Kijima H., Nakahata H., Asai S., Miyachi H., et al.: "Reversal of drug resistance using hammerhead ribozymes against multidrug resistance-associated protein and multidrug resistance 1 gene"Intern. J. Oncol.. 21. 1021-1026 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Miyachi H, Kobayashi H, Asai S, Shinbata T, Arami S and Takemura Y.: "Gene expression profiling for predicting treatment responsiveness of cancer"Recent Adv Res Updates. 3. 5-12 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Efferth T, Verdorfer I, Miyachi H, et al:: "Genomic imbalance in drug-resistant T-cell acute lymphoblastic CEM leukemia cell lines"Blood Cells Mol Dis.. 29. 1-13 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Asai S, Miyachi H, Kobayashi H, Takemura Y and Ando Y.: "A large diversity in transport-mediated methotrexate resistance in human leukemia cell line CCRF-CEM established in a high concentration of leucovorin"Cancer Science. in press. (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Miyachi H, et al.: "Gene expression profiling for predicting treatment responsiveness of cancer"Recent Adv Res Updates. 3. 5-12 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 宮地勇人, 他: "cDNAマイクロアレイ技術の臨床検査への応用.遺伝子発現プロファイル解析による癌の薬剤耐性形質の評価"臨床病理. 50. 161-168 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Nagata J., Kijima H., Nakahata H., Asai S., Miyachi H., et al.: "Reversal of drug resistance using hammerhead ribozymes against multidrug resistance-associated protein and multidrug resistance 1 gene"Intern J Oncol.. 21. 1021-1026 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Asai S, Miyachi H, Kobayashi H, Takemura Y, Ando Y.: "A large diversity in transport-mediated methotrexate resisitance in human leuekmia cell line CCRF-CEM established in a high concentration of leucovorin"Cancer Science. (in press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Takemura Y, Kobayashi H, Miyachi H: "Antifolate resistance and its circumvention by new analogues"Human Cell. 14. 185-202 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Kobayashi H, Takemura Y, Miyachi H: "Novel approaches to reversing anti-cancer drug resistance using gene-specific therapeutics"Human Cell. 14. 172-181 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 宮地勇人, 小林広幸, 浅井さとみ, 荒見真一郎, 新畑智也, 三瀬静男: "cDNAマイクロアレイ技術の臨床検査への応用:遺伝子発現プロファイル解析による癌の薬剤耐性形質の評価"臨床病理. (印刷中). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] 宮地勇人, 他: "癌の薬剤耐性とその克服-基礎と臨床-"宇宙堂. 41 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 宮地勇人 他.: "癌の薬剤耐性とその克服 基礎と臨床"宇宙堂八木書店. 119 126 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] 宮地勇人 他.: "癌の薬剤耐性とその克服 基礎と臨床"宇宙堂八木書店. 334-343 (2001)

    • Related Report
      2000 Annual Research Report

URL: 

Published: 2000-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi