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A study for cancer gene therapy through Fas-mediated apoptosis

通过 Fas 介导的细胞凋亡进行癌症基因治疗的研究

基本信息

批准号：
10671099
负责人：
TAKEDA Yasutaka
金额：
$ 2.18万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10671099/
关键词：
Cancer gene therapy Fas FasL system Apoptosis Murine hepatoma Murine mammary tumor

项目摘要

We examined the antitumor activity of anti-Fas Ab (Jo2) and soluble recombinant FasL (rFasL) against MH134 cells transfected with Fas cDNA (F6B). Original MH134 cells do not express Fas and are refractory to Fas-mediated apoptosis. In vitro, rFasL and Jo2 (200ng/ml) induced complete cell death of F6b when determined with MTT assay. This cell death was proved to be through apoptosis by Hoechist staining. On the other hand, in vivo, we investigated antitumor effects of rFasL and Jo2 on F6b using double-mutant mice (C3H-gld/gld ・ Ipr/Ipr). Both therapeutic agents induced temporary antitumor effects but did not produce complete cure of tumors in any mice. Recently, it has been reported that many tumors express Fas antigen on their cell surface. Therefore, we established C8h line by transfecting Fas cDNA to MM2 cells which constitutively express Fas at a low level but are resistent to Fas-mediated apoptosis. In vitro, C8h cells were killed through apoptosis completely with rFasL but partially with Jo2. The result suggests that rFasL and Jo2 may bind to different epitopes on Fas antigen and induce apoptosis through different signal pathways. In addition, rFasL appears to be superior to Jo2, because tumor cells developed the resistance to Fas-mediated apoptosis more easily when treated with rFasL than Jo2.

我们检测了抗Fas抗体（Jo 2）和可溶性重组FasL（rFasL）对转染Fas cDNA（F6 B）的MH 134细胞的抗肿瘤活性。原始的MH 134细胞不表达Fas，并且对Fas介导的凋亡是难治的。MTT法检测rFasL和Jo 2（200 ng/ml）诱导F6 b细胞完全死亡。Hoechist染色证实细胞死亡是通过凋亡实现的。另一方面，在体内，我们使用双突变小鼠（C3 H-gld/gld · Ipr/Ipr）研究了rFasL和Jo 2对F6 b的抗肿瘤作用。这两种治疗剂诱导暂时的抗肿瘤作用，但没有产生完全治愈肿瘤的任何小鼠。近年来，有报道称许多肿瘤细胞表面表达Fas抗原。因此，我们将Fas cDNA转染到组成型低表达Fas但对Fas介导的凋亡有抵抗力的MM 2细胞中，建立了C8 h细胞系。在体外，C8 h细胞通过凋亡完全与rFasL，但部分与乔2。结果提示rFasL和Jo 2可能分别与Fas抗原上不同的表位结合，通过不同的信号通路诱导细胞凋亡。此外，rFasL似乎是上级的乔，因为肿瘤细胞更容易发展抵抗Fas介导的凋亡时，用rFasL比乔2处理。