モノクロトール抗体,遺伝子を用いた耐性癌の基礎的な診断と治療にする研究

• Project/Area Number: 04151030
• Research Category: Grant-in-Aid for Cancer Research
• Allocation Type: Single-year Grants
• Research Institution: Mie University
• Principal Investigator: 白川 茂 三重大学, 医学部, 教授 (20026850)
• Co-Investigator(Kenkyū-buntansha): 曽根 三郎 徳島大学, 医学部附属病院, 講師 (40145024) ; 玉置 憲一 東海大学, 医学部, 教授 (50055860) ; 上田 龍三 愛知県がんセンター研究所, 化学療法部, 部長 (20142169) ; 下山 正徳 国公がんセンター中央病院, 副院長 (50110002) ; 鶴尾 隆 東京大学, 応用微生物研究所, 教授 (00012667)
• Project Period (FY): 1992
• Project Status: Completed (Fiscal Year 1992)
• Budget Amount: ¥17,000,000 (Direct Cost: ¥17,000,000); Fiscal Year 1992: ¥17,000,000 (Direct Cost: ¥17,000,000)
• Keywords: 多剤耐性(MDR)腫瘍 / P糖蛋白(P-glycoprotain;P-gp) / 抗P-gp抗体(MRK16,C219,U1C) / 多剤耐性遺伝子(MDR1) / MDR1mRNA発現 / ヒト・マウスキメラ抗体 / 免疫磁気ビーズ(MACS)法 / rhodamino123effluxtest

Research Abstract

(1)MRK-16によるP糖蛋白(P-gp)検出感度増強に免疫磁気ビーズ(MACS)法を用い,これとFACS法の組み合せで,臨床試料の耐性度診断に有用と考えられた(鶴尾)。(2)レーザー顕微鏡を用い,P-gpにより細胞内外の薬剤移動が規定された蛍光物質(adriamycin,rhadamine)と細胞を孵置し,細胞内蛍光強度や分布から多剤耐性(MDR)細胞の検出を試み,CyA誘導体PSC833の添加にて判定がより容易になった(下山)。(3)固型腫瘍のMDR1遺伝子/P-gp発現をRT-PCRと免疫染色で検討し,P-gp同定には後者が有用であるが感度が劣り,抗P-gp抗体の種類(C219,MRK16)で感度,細胞内局在に差があり,gliomoではMDR1遺伝子発現,P-gp産生は毛細血管に限局していた(玉置)。(4)正常及び悪性リンパ腫のリンパ筋につき抗P-gp抗体U1C2による免疫染色とRT-PCRによるMDR1mRNAの検討を行い,P-gp及びMDR1遺伝子発現はいずれのリンパ組識にも認められたが,P-gp陽性細胞はT-201ehistiocyteで腫瘍細胞ではなかった(白川)。(5)初診時急性白血病細胞(AML50例,ALL25例)につきP-gp/MDR1発現をRT-PCRを中心に検索し,免疫表現型と対比した。MDR1mRNAの発現はM1に高く,CD34(+)AML,CD7(+)AMLで有意に高く,幹細胞白血病に近縁なCD7(+)SCD3(-)CDU(-)CD8(-)ALLは8例中6例(75%)に発現され,予後不良であった(白川)。(6)3種類のamplimerを用いたRT-PCRでMDR1発現を検索し,いずれのamplimerでもMDR化したAML,ALL,CML-BCで33〜67%陽性,乳癌,卵巣癌とも原発巣より転移巣系で高値であった(向山)。また未分化甲状腺癌は予後最悪で,細胞株8株全例P-gp/MDR1の発現がみられた(菅原)。(7)P-gp抗体(MRK16,MRK17)を介したADCCの誘導に,NK細胞はMRK16のみ,単球は両抗体を介して如き,ヒト・マウスキメラ抗体はNK細胞をeffectorとしたADCC反応を増強させた。M-CSF遺伝子をMDR卵巣癌細胞に移入し,種々のM-CSF産生株を得,治療実験への応用を考えている(曽根,鶴尾)。(8)MDR克服にPSC833が有望であり(下山),MDR1発現の高いCD7(+)AMLでrhadamineeffluxtestが検討された(白川)。

Research Products

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