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Flow cytometric analysis of low-grade multidrug resistance using fluo-e : a comparison with rhodamine-123

Research Project

Project/Area Number 04670582
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field Pediatrics
Research InstitutionKanazawa University

Principal Investigator

KOIZUMI Shoichci  Kanazawa University School of Medicine, Associate Professor, 医学部, 助教授 (50019973)

Project Period (FY) 1992 – 1993
Project Status Completed (Fiscal Year 1993)
Budget Amount *help
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1993: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1992: ¥1,100,000 (Direct Cost: ¥1,100,000)
KeywordsMultidrug resistance / MDR1 gene / P-glycoprotein / Fluo-3 / Rhodamine-123 / K562 cells / Leukemia / Chemotherapy / K562cells / 白血病 / 化学療法 / 薬剤耐性 / P-糖蛋白 / 多剤耐性 / ベラパミル / セファランチン
Research Abstract

Six cell lines including drug-sensitive K562/parent cells, multidrug-resistant(MDR) K562/VCR, K562/ADR and K562/tetrahydropyranyl(THP)-ADR cells, methotrexate-resistant K562/MTX and revertant K562/ADR-R cell lines were used in this study : Although P-glycoprotein(P-gp)was overexpressed only in K562/VCR and K562/ADR cells, the RT-PCR method revealed distinct expression of MDR1mRNA even in the K562/parent cells. Intracellular accumulation of Fluo-3 and rhodamine-123 (Rh-123) was measured by incubation of cells with 4 muM Fluo-3 or 1muM Rh-123 for 20 min. by using a flow cytometer. Verapamil(Ver, 20 muM) or cepharanthine (biscoclaurine alkaloid, CP, 10muM) was added just before the addition of the fluorescent agents, Efflux patterns were also studied 30 and 60 min. after incubation with or without Ver ad CP.Increased intracellular accumulation and delayd efflux pattern of Fluo-3 by Ver and CP were shown in MDR K562/VCR and K562/ADR cells, and most interestingly, similar increase of uptake and delayd efflux pattern of Fluo-3 by Ver and CP butin a lower grade, were also demonstrated in P-gp-non-expressed K562/parent, K562/MTX and K562/THP cells. Accumulation of Rh-123 waas not influenced by Ver and CP.Thirteen of 31 clinical leukemic cell samples expressing MDR1mRNA, but not P-gp demonstrated enhancement of intracellular accumulation of Fluo-3 by Ver and Cp, indicating that flow cytometric functional analysis using Fluo-3 with Ver and CP may be usefl for more sensitive functional assay of low-grade multidrug resistance.

Report

(3 results)
  • 1993 Annual Research Report   Final Research Report Summary
  • 1992 Annual Research Report
  • Research Products

    (16 results)

All Other

All Publications (16 results)

  • [Publications] Koizumi,S.: "Rapid and sensitive detection of low-grade multidrug resistance(MDR)in leukemic cells using the fluorescent indicator,Fluo-3." Proc.Am.Assoc.Cancer Res.33. 481-481 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 小泉晶一: "蛍光プローベFluo-3を用いた、低レベル多剤耐性細胞の迅速解析" Proc.Jpn.Cancer Assoc.(日本癌学会総会記事). 51. 418-418 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Koizumi,S.: "Efficacy of intermittent methotrexate(MTX)and 6-mercaptopurine(6MP)in maintenance chemotherapy for childhood acute lymphoblastic leukemia(ALL)." Proc.Am.Soc.Clin.Oncol.11. 286-286 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 小泉晶一: "多剤耐性細胞におけるP-糖蛋白発現と蛍光プローベFluo-3の細胞内蓄積の良い相関性" Proc.Jpn.Cancer Assoc.(日本癌学会総会記事). 52. 638-638 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 小泉晶一: "蛍光プローベFluo-3を用いた多剤耐性細胞の高感度迅速解析" 医学のあゆみ. 165. 131-132 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Koizumi,S.: "Improvement in treatment of childhood acute lymphoblastic leukemia:a 10-year study by the Children's Cancer and Leukemia Study Group." Intern.J.Hematol.59(in press). (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Koizumi, S.: "Rapid and sensitive detection of low-grade multidrug resistance (MDR) in leukemic cells using the fluorescent indicator, Fluo-3" Proc. Am. Assoc. cancer Res.33. 481 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Koizumi, S.: "Rapid and sensitive detection of low-grade multidrug resistance (MDR) in leukemic cells using the fluorescent indicator, Fluo-3" Proc. Jpn. Cancer Assoc.51. 418 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Koizumi, S.: "Efficacy of intermitent methotrexate (MTX) and 6-mercaptopurine (6MP) in maintenance chemotherapy for childhood acute lymphoblastic leukemia (ALL)" Proc. Am. Soc. Clin. Oncol.11. 286 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Koizumi, S.: "Good correlation between expression of P-glycoprotein and accumulation of fluorescent Fluo-3 probe." Pro. Jpn. cancer Assoc.52. 638 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Koizumi, S.: "A rapid and sensitive assessment ofmultidrug resistance using an Fluo-3 fluorescent phobe" J.clin. Exp. Med. (IGAKU NO AYUMI). 165. 131-132 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Koizumi, S.: "Improvement in treatment of childhood acute lymphoblastic leukemia : a 10-year study by the Children's Cancer and Leukemia Study Group" Intern. J.Hematol.(in press). (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Katayama, K.: "Successful peritransplant therapy in children with active hepatosplenic candidiasis" Intern. J.Hematol.(in press). (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 小泉 晶一: "蛍光プローベFluo-3を用いた多剤耐性細胞の高感度迅速解析" 医学のあゆみ. 165. (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] Koizumi S.: "Enzyme studies of methotrexate-resistant human leukemic cell(K562) cell subclones." Leukemia Reasearch. 16. 565-569 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 小泉 晶一: "小児急性リンパ性白血病の治療研究:CCLSG-L874,I874プロトコールの成績" 日本小児血液学会雑誌. 6. 446-454 (1992)

    • Related Report
      1992 Annual Research Report

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Published: 1992-04-01   Modified: 2016-04-21  

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