Shiraishi Tetsuya, Suzuyama Kenji, Okamoto Hiroaki, Mineta Toshihiro, Tabuchi Kazuo, Nakayama Kazuyuki, Shimizu Yusuke, Tohma Junko, Ogihara Takuo, Naba Hiroyasu, Mochizuki Hidenori, Nagata Shigekazu
Department of Neurosurgery, Faculty of Medicine, Saga University, Japan.
Biochem Biophys Res Commun. 2004 Sep 10;322(1):197-202. doi: 10.1016/j.bbrc.2004.07.098.
Fas (CD95) ligand (FasL) has the ability to induce apoptosis in Fas-expressing glioma cells by binding to Fas. Several molecular species have been designed to be soluble Fas ligands for therapeutic purposes. We successfully constructed a chimeric soluble FasL by fusing an isoleucine zipper motif for self-oligomerization and a FLAG sequence to the extracellular domain of the human Fas ligand (FIZ-shFasL). The cytotoxic effect of FIZ-shFasL on Jurkat cells was equivalent to that of membrane-bound FasL and approximately 10-fold stronger than that of agonistic anti-Fas antibody (CH-11). Flow cytometric analysis demonstrated that the differential Fas expression of human brain tumor cell lines partially correlated with levels of apoptosis through FIZ-shFasL. The upper limit of FIZ-shFasL for safe systemic administration to rat is estimated as below 2 microg/ml in plasma concentration. FIZ-shFasL could be applicable as a therapeutic agent for cancer.
Fas（CD95）配体（FasL）能够通过与Fas结合，诱导表达Fas的胶质瘤细胞发生凋亡。为了治疗目的，已经设计了几种分子形式的可溶性Fas配体。我们通过将用于自我寡聚化的异亮氨酸拉链基序和一个FLAG序列融合到人Fas配体（FIZ-shFasL）的胞外域，成功构建了一种嵌合可溶性FasL。FIZ-shFasL对Jurkat细胞的细胞毒性作用与膜结合FasL相当，且比激动性抗Fas抗体（CH-11）强约10倍。流式细胞术分析表明，人脑肿瘤细胞系中Fas表达的差异与通过FIZ-shFasL诱导的凋亡水平部分相关。FIZ-shFasL对大鼠进行安全全身给药时，血浆浓度的上限估计低于2微克/毫升。FIZ-shFasL有望作为一种癌症治疗药物。
