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

利用野百合醇抗体和基因进行耐药性癌症的基础诊断和治疗研究

• 批准号: 04151030
• 负责人: 白川 茂
• 金额: $ 10.88万
• 依托单位: Mie University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Cancer Research
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-04151030/
• 关键词: 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

(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)。

(1)MRK-16 P-glycoprotein (P-gp) detection sensitivity increased by the use of immunomagnetic staining (MACS) method, and FACS method was used in combination, clinical samples of resistance diagnosis useful test. (2)P-gp regulation of intracellular and extracellular drug movement and regulation of adriamycin,rhadamine and cell incubation, intracellular light intensity and distribution, detection of MDR cells, and determination of addition of CyA inducer PSC833 are easy to detect. (3)RT-PCR and immunostaining assay for MDR1 gene/P-gp expression in solid tumors,P-gp identification and sensitivity to the latter, sensitivity to anti-P-gp antibody types (C219, MRK16), intracellular differences,gliomo and MDR1 gene expression,P-gp production and capillary limitation (Yuzhi). (4)P-gp and MDR1mRNA were detected by immunostaining and RT-PCR in normal and abnormal cells, and T-201 histiocyte in P-gp positive cells and tumor cells (Shirakawa). (5)At the time of initial diagnosis, acute leukemia cells (AML50 cases,ALL25 cases) were detected for P-gp/MDR1 expression, RT-PCR was used for central detection, and immunophenotype was compared. MDR1 mRNA expression was high in M1, CD34 (+)AML, CD7 (+)AML was intentionally high, stem cell leukemia was close to CD7 (+) SCD3 (-)CDU(-) CD8 (-)ALL, and was detected in 6 of 8 cases (75%). (6)3 RT-PCR detection of MDR1 in the presence of amplimer, MDR AML,ALL,CML-BC 33 ~ 67% positive, breast cancer, ovarian cancer, MDR1 gene detection, migration system high value (Xiang Shan). 8 undifferentiated thyroid carcinoma cell lines showed P-gp/MDR1 expression in all cases. (7)P-gp antibodies (MRK16, MRK17) mediate ADCC induction,NK cells respond to MRK16 and CD4 + antibodies mediate ADCC induction, CD4 + antibodies respond to NK cells respond to MRK16 and CD4 + antibodies respond to ADCC induction. M-CSF gene was transplanted into MDR ovarian cancer cells, and M-CSF producing strains were obtained. (8)MDR overcomes PSC833 and is expected to be detected (downhill),MDR1 appears high in CD7 (+)AML and rhadamineeffluxtest is detected (Shirakawa).

项目成果

Sygawara I,et al: "Proparation and charactnization of a murine mono-clonal antibody (MDR3M) reactive with maru gene product." Jpn J Cancer Res. 83. 795-797 (1992)

Sygarara I 等人：“与 maru 基因产物反应的鼠单克隆抗体 (MDR3M) 的制备和表征。”

Adachi M,et al: "Protain-tyrosine phosphatase expression in pre-B cell NALM-6" Cancer Res. 52. 737-740 (1992)

Adachi M 等人：“前 B 细胞 NALM-6 中的蛋白质酪氨酸磷酸酶表达”Cancer Res。

Nakase K,et al: "Diagnostic and climical importance of interleubin-2 veceptor algha chain expression on non-T-coll acute loukemda cells." Br J Idaemaeol. 80. 317-326 (1992)

Nakase K 等人：“非 T-coll 急性 loukemda 细胞上白细胞介素 2 载体藻链表达的诊断和临床重要性。”

Ishii S,et al: "Expression of 85-kDa protein of adriamycin-resistant tunon cells during hematopoietic diffenntiation of THP-1 and MEL cells." Jpn J Cancer Res. 83. 107-112 (1992)

Ishii S 等人：“THP-1 和 MEL 细胞的造血分化过程中阿霉素耐药 Tunon 细胞的 85 kDa 蛋白的表达”。

島田 安博,下山 正徳: "MDR遺伝子と薬剤耐性の克服" 最新医学. 48. 243-249 (1993)

Yasuhiro Shimada，Masanori Shimoyama：“MDR 基因与克服耐药性”现代医学 48. 243-249 (1993)。