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Effects of specific antisense oligonucleotide inhibition of Fas expression on T cell apoptosis induced by Fas ligand.

机译:反义寡核苷酸特异性抑制Fas表达对Fas配体诱导的T细胞凋亡的影响。

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摘要

To investigate the effect of specific antisense oligodeoxynucleotide (ASODN) inhibition of Fas expression on T cell apoptosis induced by hepatocarcinoma cells. Fas receptor (Fas) and Fas ligand (FasL) expressed by the hepatocarcinoma cell line HepG2.2.15 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 HepG2.2.15 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. It was found that HepG2.2.15 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 hepatocarcinoma cells were all decreased. As a conclusion, it is evident that hepatocarcinoma 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 hepatocarcinoma cells.
机译:研究特异性反义寡聚脱氧核苷酸(ASODN)抑制Fas表达对肝癌细胞诱导的T细胞凋亡的影响。通过流式细胞术(FCM)检测肝癌细胞系HepG2.2.15和Jurkat T细胞表达的Fas受体(Fas)和FasL配体(FasL),并通过共培养法检测FasL诱导T细胞凋亡的能力。用HepG2.2.15细胞和Jurkat T细胞体外培养。通过脂质转染法将Fas-ASODN转染Jurkat细胞,并分别通过RT-PCR,FCM和共培养法研究Fas-ASODN对Fas mRNA水平,T细胞表面Fas表达和细胞凋亡的影响。如共培养测定所证实,发现表达功能性FasL的HepG2.2.15细胞可诱导Jurkat细胞的凋亡。 Fas ASODN转染Jurkat细胞后,肝癌细胞诱导的Fas mRNA表达水平,Fas表达率和凋亡率均降低。结论是,很明显,表达FasL的肝癌细胞可以诱导表达Fas的T细胞凋亡,这表明Fas ASODN的转染可以部分转化肝癌细胞诱导的免疫抑制条件。

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