{"created":"2023-07-27T06:51:19.659448+00:00","id":44710,"links":{},"metadata":{"_buckets":{"deposit":"5cc28ad0-a4e6-4a79-900e-82b5b49ef776"},"_deposit":{"created_by":18,"id":"44710","owners":[18],"pid":{"revision_id":0,"type":"depid","value":"44710"},"status":"published"},"_oai":{"id":"oai:kanazawa-u.repo.nii.ac.jp:00044710","sets":["2812:2813:2840"]},"author_link":["77002","52683"],"item_9_biblio_info_8":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"1994-03","bibliographicIssueDateType":"Issued"},"bibliographicPageStart":"6p.","bibliographicVolumeNumber":"1992-1993","bibliographic_titles":[{"bibliographic_title":"平成5(1993)年度 科学研究費補助金 一般研究(C) 研究成果報告書"},{"bibliographic_title":"1993 Fiscal Year Final Research Report","bibliographic_titleLang":"en"}]}]},"item_9_creator_33":{"attribute_name":"著者別表示","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{}],"nameIdentifiers":[{},{}]}]},"item_9_description_21":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"[目的]P-糖蛋白のefflux pumpとしての機能性を応用して、多剤耐性(MDR)細胞の、新しい蛍光プローベFluo-3を用いた機能的解析の可能性を検討し、Rhodamine-123と比較した。[方法]薬剤感受性K562親株、多剤耐性K562/VCR、K562/ADR細胞、その感受性復元細胞、MTXのみ耐性で非多剤耐性K562/MTX細胞、多剤耐性であるがP-糖蛋白陰性K562/THPADR細胞を使用した。細胞(10^6/ml 5%FCS加RPMI1640)を37℃保温後、Fluo-3A/M(4μM)またはRh-123(1μM)を添加した。20分後に530nmで平均蛍光強度(mcf)をflow cytometer(Cytoron Absolute)で測定した。さらにP-糖蛋白阻害剤のverapamil(Ver,20μM)やcepharanthine(CP,10μM)を添加してmcfの変化を観察した。Effluxパターンは細胞を洗浄後30分、60分で検討した。P-糖蛋白発現はMRK16抗体を用いた免疫蛍光染色法によった。MDRmRNAはRT-PCR法によった。[結果]K562/VCRとK562/ADRではFluo-3,Rh-123ともに細胞内蓄積低下とVer/CPによる排泄遅延がみられた。P-糖蛋白発現のみられないK562/THPADR,K562/MTXではVer/CPによるFluo-3の排泄遅延が認められたがRh-123ではみられなかった。MDR1mRNA発現はP-糖蛋白陰性の細胞も含めてすべての細胞でみられた。[結論]以上より、Rh-123はP-糖蛋白発現をより明確に表現したが、Fluo-3はMDR1mRNAを発現する低レベルの多剤耐性の検出に有用かもしれない。","subitem_description_type":"Abstract"},{"subitem_description":"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.","subitem_description_type":"Abstract"}]},"item_9_description_22":{"attribute_name":"内容記述","attribute_value_mlt":[{"subitem_description":"研究課題/領域番号04670582, 研究期間(年度):1992-1993","subitem_description_type":"Other"},{"subitem_description":"出典：「Fluo-3を用いた、低レベルの抗癌剤多剤耐性細胞の迅速解析と耐性克服剤の検討」研究成果報告書　課題番号04670582\n(KAKEN：科学研究費助成事業データベース（国立情報学研究所）)\n　　　本文データは著者版報告書より作成","subitem_description_type":"Other"}]},"item_9_identifier_registration":{"attribute_name":"ID登録","attribute_value_mlt":[{"subitem_identifier_reg_text":"10.24517/00051051","subitem_identifier_reg_type":"JaLC"}]},"item_9_publisher_17":{"attribute_name":"公開者","attribute_value_mlt":[{"subitem_publisher":"金沢大学医薬保健研究域医学系"}]},"item_9_relation_28":{"attribute_name":"関連URI","attribute_value_mlt":[{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/search/?qm=50019973"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/search/?qm=50019973","subitem_relation_type_select":"URI"}},{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-04670582/"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-04670582/","subitem_relation_type_select":"URI"}},{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/report/KAKENHI-PROJECT-04670582/046705821993kenkyu_seika_hokoku_gaiyo/"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/report/KAKENHI-PROJECT-04670582/046705821993kenkyu_seika_hokoku_gaiyo/","subitem_relation_type_select":"URI"}}]},"item_9_version_type_25":{"attribute_name":"著者版フラグ","attribute_value_mlt":[{"subitem_version_resource":"http://purl.org/coar/version/c_ab4af688f83e57aa","subitem_version_type":"AM"}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2018-06-14"}],"displaytype":"detail","filename":"ME-PR-KOIZUMI-S-kaken 1994-6p.pdf","filesize":[{"value":"271.8 kB"}],"format":"application/pdf","licensetype":"license_11","mimetype":"application/pdf","url":{"label":"ME-PR-KOIZUMI-S-kaken 1994-6p.pdf","url":"https://kanazawa-u.repo.nii.ac.jp/record/44710/files/ME-PR-KOIZUMI-S-kaken 1994-6p.pdf"},"version_id":"966e1ec1-68e4-4447-870d-ef5bda665c64"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"jpn"}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"research report","resourceuri":"http://purl.org/coar/resource_type/c_18ws"}]},"item_title":"Fluo-3を用いた、低レベルの抗癌剤多剤耐性細胞の迅速解析と耐性克服剤の検討","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"Fluo-3を用いた、低レベルの抗癌剤多剤耐性細胞の迅速解析と耐性克服剤の検討"},{"subitem_title":"Flow cytometric analysis of low-grade multidrug resistance using fluo-e : a comparison with rhodamine-123","subitem_title_language":"en"}]},"item_type_id":"9","owner":"18","path":["2840"],"pubdate":{"attribute_name":"公開日","attribute_value":"2018-06-14"},"publish_date":"2018-06-14","publish_status":"0","recid":"44710","relation_version_is_last":true,"title":["Fluo-3を用いた、低レベルの抗癌剤多剤耐性細胞の迅速解析と耐性克服剤の検討"],"weko_creator_id":"18","weko_shared_id":-1},"updated":"2023-07-27T14:46:49.888792+00:00"}