新規抗癌剤開発のためのリピッドラフト解析

脂筏分析用于开发新的抗癌药物

• 批准号: 18659287
• 负责人: 梅原 久範
• 金额: $ 2.05万
• 依托单位: Kanazawa Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Exploratory Research
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-18659287/
• 关键词: リピッドラフト; アポトーシス; スフィンゴミエリン

1)スフィンゴミエリン欠失細胞株細胞株におけるFas誘導apoptosisの解析:スフィンゴミエリン欠質細胞およびコントロール細胞を抗Fas抗体(CH11)で刺激し,apoptosisをsub G1法でFACSで解析し,両細胞におけるapoptosisの差異を明らかにした.Fas架橋刺激による,DISC形成の検出,Caspase活性の測定,ミトコンドリア膜電位の測定を行なった.2)細胞膜lipid raftのconfocal microscopyによる解析:スフィンゴミエリン欠失細胞株におけるlipid raftの形態および機能を明らかにした.細胞膜でのスフィンゴミエリンの局在を明らかにするために,スフィンゴミエリンに特異的に結合するMBP結合1yseninとPE標識抗MBP抗体を用いて染色した.Fas架橋刺激によるCluster形成をFITC-,PE-標識Fas抗体で検出し,ラフト構成におけるganglioside GM1とスフィンゴミエリンおよびFasの動態を明らかにした.スフィンゴミエリン欠失細胞株では,Fasの凝集が抑制されていた.3)Fas誘導シグナルの差異の検討:Fas架橋刺激に伴う,FADD,Caspase-8とFasとの結合をwestern blottingおよび免疫沈降法を用いて検討した.リピッドラフトをsucrose gradient法により分画し,ganglioside GM1量とスフィンゴミエリン量を比較検討した.Fas刺激に伴うFADDやCaspaseなどのシグナル伝達物質のlipidraftへの移行を検討した.スフィンゴミエリン欠失細胞では,Fas刺激によるcaspase-8,caspase-3活性が抑制されていた.また,Fasに会合するFADD,Caspase-8量も低下していた.その結果Fasによるアポトーシスがスフィンゴミエリン欠失細胞では障害されていることが明らかになった.

1) Analysis of Fas induced apoptosis in cell lines: Fas induced apoptosis induced by anti-Fas antibody (CH11) stimulated by Fas bridging stimulation, apoptosis induced by sub-G1 FACS analysis, differences in apoptosis induced by anti-Fas antibody (CH11) stimulated by Fas bridging stimulation,DISC formation, determination of Caspase activity, measurement of Caspase membrane potential; 2) analysis of lipid membrane by confocal microscopy: Fas induced apoptosis induced by anti-Fas antibody stimulated by sub-G1 FACS analysis, differences in apoptosis induced by Fas bridging stimulation, and changes in lipid membrane potential induced by Fas bridging stimulation. Cell membrane binding to MBP and PE-labeled anti-MBP antibody was detected by FITC-, PE-labeled anti-Fas antibody staining.Fas bridge stimulation was used to detect Cluster formation and Fas ganglioside GM1. 3)Fas induction and differentiation:Fas bridging stimulation,FADD,Caspase-8 and Fas binding, western blotting and immunoprecipitation. Fas stimulation is accompanied by Caspase FADD and the migration of lipid in the target substance is studied. Fas stimulation inhibited caspase-8 and caspase-3 activity in the absence of Fas. FADD,Caspase-8 levels are low. As a result, the Fas family was divided into two groups: the first group was divided into three groups: the second group was divided into three groups: the first group was divided into three groups: the first group was divided into three groups: the second group was divided into three groups: the first group was divided into three groups: the first group was divided into three groups: the second group was divided into three groups: the first group was divided into three groups: the second group was divided into three groups: the first group was divided into three groups: the first group was divided into three groups: the second group was divided into three groups: the first group was divided into three groups: the first group was divided into three groups: the second group was divided into three groups: the first group was divided into three groups: the first group was divided into three groups; the second group was divided into three groups: the third group was divided into three groups; the third group was divided into three groups: the first group was divided into three groups; the third group was divided into three groups; the third group was divided into three groups; the third

项目成果

Anti-aminoacyl-tRNA synthetase antibodies in clinical course prediction of interstitial lung disease complicated with idiopathic inflammatory myopathies

• DOI: 10.1080/08916930600622884
• 发表时间: 2006-05-01
• 期刊: AUTOIMMUNITY
• 影响因子: 3.5
• 作者: Yoshifuji, H;Fujii, T;Mimori, T
• 通讯作者: Mimori, T

Successful salvage therapy of continuous low-dose irinotecan for a patient with relapsed and refractory diffuse large B-cell lymphoma

连续小剂量伊立替康成功挽救复发难治性弥漫性大 B 细胞淋巴瘤患者

• 发表时间: 2006
• 期刊: J.Rinsho Retsueki 46
• 作者: Masaki Y;Sawaki T;Shimoyama;Umehara H. et al.
• 通讯作者: Umehara H. et al.

Sjogren's syndrome and lymphoma development.(Review)

干燥综合征和淋巴瘤的发展。（综述）

• 发表时间: 2007
• 期刊: Clin. Immunol. Rev. 3
• 作者: Dong;L.,
• 通讯作者: L.,

Detection of serum hepcidin in renal failure and inflammation by using ProteinChip System

• DOI: 10.1182/blood-2005-10-4043
• 发表时间: 2006-08-15
• 期刊: BLOOD
• 影响因子: 20.3
• 作者: Tomosugi, Naohisa;Kawabata, Hiroshi;Ishikawa, Isao
• 通讯作者: Ishikawa, Isao

Efficacy and safety of mizoribine for the treatment of Sjogren's syndrome : a multicenter open-label clinical trial

米佐立滨治疗干燥综合征的有效性和安全性：多中心开放临床试验

• 发表时间: 2007
• 期刊: Mod Rheumatol 17(6)
• 作者: Nakayamada S;Saito K;Umehara H;Ogawa N;Sumida T;Ito S;Minota S;Nara H;Kondo H;Okada J;Mimori T;Yoshifuji H;Sano H;Hashimoto N;Sugai S;Tanaka Y
• 通讯作者: Tanaka Y