FasL反义寡核苷酸逆转肝癌细胞免疫逃逸的实验研究  

The FasL antisense oligonucleotide could reverse the immune escape in hepatocarcinoma cell line

作　　者：张娇[1] 刘倩[1] 王杰[2] 毛海婷[2] 

机构地区：[1]山东大学临床医学院山东省千佛山医院消化科,山东济南250014 [2]山东省医学科学院基础医学研究所,山东济南250062

出　　处：《中国现代普通外科进展》2006年第3期147-150,共4页Chinese Journal of Current Advances in General Surgery

基　　金：山东省自然科学基金资助项目(Y2002C45)

摘　　要：目的:研究FasL反义寡核苷酸对肝癌细胞Fas/FasL途径免疫逃逸的逆转作用。方法:用流式细胞术检测肝癌细胞系HepG2.2.15细胞表面Fas、FasL的表达及Jurkat细胞表面Fas的表达;用HepG2.2.15细胞与Jurkat细胞共培养方法,研究FasL诱导T细胞凋亡的功能;用脂质体介导法转导FasL反义寡核苷酸入肝癌细胞,并通过检测细胞表面FasL表达、RTPCR及细胞共培养方法研究FasL反义寡核苷酸转导对肝癌细胞免疫抑制的逆转作用。结果:①HepG2.2.15细胞低表达Fas,却表达有功能的FasL,与高表达Fas的Jurkat细胞共培养可诱导后者凋亡;②肝癌细胞转入FasL反义寡核苷酸后,FasLmRNA降低;细胞表面FasL表达下降;与Jurkat细胞共培养,Jurkat细胞的凋亡率下降。结论:肝癌细胞表达的FasL可抑制T细胞的免疫功能,是肝癌细胞免疫逃逸的重要机制之一;FasL反义寡核苷酸转导可以逆转肝癌细胞的免疫抑制作用。Objective： To Investigate the reverse action of FasL antisense ollgonucleotide on the Immune escape through Fas/FasL pathway in hePatocarcinoma cells.Methods ： Fas, FasL expression of hePatocarclnoma cell line HePG2.2.15 and Fas expression of Jurkat cell were examined bY flow cytometry. FasL function of inducing T-lymphooytes to apoptosls was assessed by cocultrue assays in vitro using hepatocarcinoma cells HepG2.2.15 and Ourkat cells. The hepatocarcinoma cells were transfected with FasL antisense ollgonucleotlde mediated by llpofectin.The reverse action was investlsated by the ways of detecting the FasL expresslon,RT-PCR and coculture assays. Mesults： （1）HepG2.2.15 cells express Fas at a very low level, but express functional FasL which could Induce Jurkat Cells with high Fas expression to apoptosis in cocultrue assays,（2）After the hepatocercinoma cells were transfected with FasL antisense ollgonucleotide,the FasL mRNA,the rate of FasL expression and the apoptotic rate of Jurkat cells in coculture assays all decreased. Conolusion：FasL expressed by the hepatocercinoma cells ooutd inhibit the Immune function of T-lymphocytes. It was one of the important mechanisms of Immune escape In hepatocarcinoma cells. FasL antisense ollgonucieotide could reverse the action of immune Inhibition.

关 键 词：肝肿瘤 FAS/FASL 免疫逃逸 寡核苷酸类 反义 

分 类 号：R735.7[医药卫生—肿瘤]