反义寡核苷酸Fas抑制T细胞凋亡

Shi-ying Zheng, D. Jiang, J. Ge, Zhengjie Shen
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引用次数: 0

摘要

研究了特异性反义寡脱氧核苷酸(ASO DN)抑制Fas表达对胃癌细胞诱导的T细胞凋亡的影响。采用流式细胞术检测胃癌细胞系SGC-7901和Jurkat T细胞表达的Fas受体(Fas)和Fas配体(FasL),并与SGC-7901细胞和Jurkat T细胞体外共培养实验检测FasL诱导T细胞凋亡的能力。用脂质体素转染Jurkat细胞Fas- asodn,分别采用RT-PCR、流式细胞术和共培养法观察Fas- asodn对Jurkat细胞Fas mRNA水平、T细胞表面Fas表达及凋亡的影响。共培养实验表明,表达功能性FasL的SGC-7901细胞可诱导Jurkat细胞凋亡。Fas ASODN转染Jurkat细胞后,Fas mRNA水平、Fas表达率及胃癌细胞凋亡率均降低。表达FasL的胃癌细胞可诱导表达FasL的T细胞凋亡,说明转染Fas ASODN可部分转化胃癌细胞诱导的免疫抑制状态。Fas ASODN可能成为肿瘤免疫治疗的重要工具。
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T Cell Apoptosis Was Inhibited by Fas of Antisense Oligonucleotide
We investigate the effect of specific antisense oligodeoxynucleotide (ASO DN) inhibition of Fas expression on T cell apoptosis induced by gastric carcinoma cell . Fas receptor (Fas) and Fas ligand (FasL) expressed by the gastric carcinoma cell line SGC-7901 and Jurkat T cells were detected by flow cytometry (FCM) and the ability of FasL-inducing T cell apoptosis was tested by co-culture assay in vitro with SGC-7901 cells and Jurkat T cells. The Jurkat cells were transfected with Fas-ASODN using lipofectin, and the effects of Fas-ASODN on Fas mRNA level, Fas expression on T cells surface, and apoptosis were investigated by RT-PCR, FCM and co-culture assay, respectively. SGC-7901 cells expressing functional FasL could induce the apoptosis of Jurkat cells as demonstrated by co-culture assays. After the Jurkat cells were transfected with Fas ASODN, the level of Fas mRNA, the expression rate of Fas and the apoptotic rate induced by gastric carcinoma cells were all decreased. Gastric carcinoma cells expressing FasL can induce apoptosis in Fas-expressing T cells, indicating that transfection of Fas ASODN can partially convert the immune inhibitory condition induced by gastric carcinoma cells. Fas ASODN may be a useful tool in the armamentarium of tumor immune therapy.
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