Induction of apoptosis in human glioma cells by recombinant human Fas lignd expressed in Baculovirus system

杆状病毒系统表达的重组人Fas配体诱导人胶质瘤细胞凋亡

• 批准号: 08671589
• 负责人: ABE Masamitsu
• 金额: $ 1.41万
• 依托单位: SAGA MEDICAL SCHOOL
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08671589/
• 关键词: Fas ligand; Fas; apoptosis; brain tumor; glioma; 脳腫瘍; アポトーシス; TUNEL; OK-432

The Fas/APO-1/CD95 ligand (FasL) is a 40kD type II transmembrane glycoprotein belonging to the tumor necrosis factor (TNF) family and has ability to induce apoptosis in sensitive target cells by binding to its receptor Fas/APO-1/CD95. Aurographa california nuclear polyhedrosis virus (AcMNPV) is the prototype virus of the family Baculoviridae. Baculovirus has polyhedrin promoter, which can express foreign proteins at levels ranging from 1 to 500 mg/liter. To develop an expression system yielding large amount of FasL,we have constructed baculovirus vector containing cDNA of FasL under polyhedrin promoter control. Hence, the expression of recombinant human FasL (rh-FasL) was detected by immunocytochemistry and Western blotting respectively. The rh-FasL showed cytotoxic activity against human glioma cell line T98G in a dose-dependent manner. The induction of apoptosis by the Fas/FasL system could be a new strategy for the treatment of malignant brain tumors, which are resistant for other conventional therapies. The baculovirus exptession system is a powerful tool to produce a large amount of biologically active rh-FasL.

Fas/APO-1/CD95配体（FasL）是一种40kD II型跨膜糖蛋白，属于肿瘤坏死因子（TNF）家族，能够通过与其受体Fas/APO-1/CD95结合，诱导敏感靶细胞凋亡。加利福尼亚核多角体病毒（AcMNPV）是杆状病毒科的原型病毒。杆状病毒具有多面蛋白启动子，可以表达1至500毫克/升的外源蛋白。为了构建高表达FasL的杆状病毒载体，在多面蛋白启动子控制下，构建了含有FasL cDNA的杆状病毒载体。因此，分别用免疫细胞化学和Western blotting检测重组人FasL （rh-FasL）的表达。rh-FasL对人胶质瘤细胞株T98G具有剂量依赖性的细胞毒活性。Fas/FasL系统诱导细胞凋亡可能是治疗恶性脑肿瘤的新策略，而恶性脑肿瘤对其他常规疗法具有耐药性。杆状病毒表达系统是产生大量具有生物活性的rh-FasL的有力工具。

项目成果

田渕和雄: "脳腫瘍をめぐる分子生物学的知見." 週刊医学界新聞(脳腫瘍-発生要因から遺伝子治療まで). 5-7 (1997)

Kazuo Tabuchi：“脑肿瘤的分子生物学发现。”周刊 Igaku Kai Shimbun（脑肿瘤 - 从发育因素到基因治疗）5-7（1997）。

田渕 和雄: "グリオーマ" 三輪書店, 162 (1997)

田渊一夫：《神经胶质瘤》三轮书店，162 (1997)

Toda K,et al.: "The cytocidal actiity of OK-432-activated mononuclear cells against human glioma cells is partly mediated through the Fas Ligand/Fas system" Jpn J Cancer Res. vol.87. 972-976 (1996)

Toda K 等人：“OK-432 激活的单核细胞对人神经胶质瘤细胞的杀细胞活性部分是通过 Fas 配体/Fas 系统介导的”Jpn J Cancer Res。

川口 正二郎: "OK-MCs(OK-432-activated mononuclear cells)の抗脳腫瘍効果におけるFas ligand/Fas系の関与." 神経化学. 35. 462-463 (1996)

Shojiro Kawaguchi：“Fas 配体/Fas 系统参与 OK-MC（OK-432 激活的单核细胞）的抗脑肿瘤作用。神经化学”。

Tabuchi K,et al.: "Expression of apoptosis-related gene productsin human brain tumors and apoptosis-inducing therapy" Nihon Rinsho. vol.54. 1922-1928 (1996)

Tabuchi K,et al.：“人脑肿瘤中凋亡相关基因产物的表达和凋亡诱导疗法”Nihon Rinsho。