特异性ASODN抑制Fas表达降低FasL介导细胞凋亡的研究

Inhibitory effect of the specific antisense oligodeoxynucleotide on Fas expression and T cell apoptosis induced by FasL

为了研究特异性Fas反义寡核苷酸(ASODN)对T细胞Fas表达及肝癌细胞诱导其凋亡的抑制作用,用脂质体介导法将Fas ASODN导入Jurkat T细胞,并通过用流式细胞术、RT-PCR及与肝癌细胞共培养方法研究Fas ASODN对T细胞Fas表达、Fas mRNA水平及凋亡的影响。结果显示:①Hep G2.2.15细胞表达有功能的FasL,可诱导Jurkat细胞凋亡;②Jurkat细胞转染Fas ASODN后,Fas mRNA降低;细胞表面Fas表达下降;与Hep G2.2.15细胞共培养后的凋亡率下降。表明Fas ASODN转染可以部分逆转肝癌细胞对T细胞的凋亡诱导作用。

To investigate the inhibitory effect of the specific antisense oligodeoxynucleotide （ASODN） on Fas expression and T cell apoptosis induced by hepatocarcinoma cells, Fasl. expressed by hepatocarcinoma cell line HepG2. 2. 15 was detected by flow cytometry（FCM） and the function of the FasL-inducing T cell apoptosis was assayed by co-culture assay in vitro with HepG2. 2. 15 cells and Jurkat cells. The Jurkat cells were trasfected with Fas-ASODN through lipofectin. Meanwhile, the influence of Fas-ASODN on the Fas expression on T cells, Fas mRNA level and apoptosis was investigated through FCM, RT-PCR ancl co-cultures. It was found that HepG2.2. 15 cells expressing the functional FasL could induce the apoptosis of Jurkat cells as demonstrated by co-culture assay, and the level of Fas mRNA, the rate of Fas expression and the apoptotic rate induced by hepatocarcinoma cell line were all decreased. As a conclusion, it is evident that hepatocarcinoma cells expressing FasL can induce the apoptosis of T cells, indicating that trasfection of Fas ASODN can partially convert the inhibitory effect induced hy heptocarcinoma cells.