FasL反义寡核苷酸逆转肝癌细胞免疫逃逸的实验研究.doc

FasL反义寡核苷酸逆转肝癌细胞免疫逃逸的实验研究 目录 TOC \o 1-9 \h \z \u 目录 1 正文 1 文1：FasL反义寡核苷酸逆转肝癌细胞免疫逃逸的实验研究 1 1 材料和方法 2 2 结 果 5 3 讨 论 6 文2：FasL反义寡核苷酸逆转肝癌细胞免疫逃逸的实验研究医学论文 7 1 材料和方法 8 2 结 果 11 3 讨 论 12 参考文摘引言： 13 原创性声明（模板） 15 文章致谢（模板） 15 正文 FasL反义寡核苷酸逆转肝癌细胞免疫逃逸的实验研究 文1：FasL反义寡核苷酸逆转肝癌细胞免疫逃逸的实验研究 The FasL antisee oligonucleotide could revee the immune escape in hepatocarcinoma cell line 【ABSTRACT】Objective:To investigate the revee action of FasL antisee oligonucleotide on the immune escape through Fas/FasL pathway in hepatocarcinoma :Fas、FasL expression of hepatocarcinoma cell line and Fas expression of Jurkat cell were examined by flow function of inducing Tlymphocytes to apoptosis was assessed by cocultrue assays in vitro using hepatocarcinoma cells and Jurkat hepatocarcinoma cells were trafected with FasL antisee oligonucleotide mediated by revee action was investigated by the ways of detecting the FasL expression，RTPCR and coculture :① 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，②After the hepatocarcinoma cells were trafected with FasL antisee oligonucleotide，the FasL mRNA，the rate of FasL expression and the apoptotic rate of Jurkat cells in coculture assays all :FasL expressed by the hepatocarcinoma cells could inhibit the immune function of was one of the important mechanisms of immune escape in hepatocarcinoma antisee oligonucleotide could revee the action of immune inhibition。 【KEY WORDS】Liver neoplasms?Fas/FasL?Immune escape?Oligonucleotide，antisee　　近年研究发现，某些肿瘤细胞表达FasL，可以与激活的T细胞表面的Fas结合，诱导T细胞凋亡，从而反击宿主免疫系统，逃避免疫监视。，探讨肝癌细胞通过Fas/FasL途径的免疫逃逸机制，并设计合成特异的FasL反义寡核苷酸（ASODN），封闭肝癌细胞FasL的表达，以期阻止免疫细胞凋亡，逆转肿瘤细胞的免疫逃逸。 1 材料和方法 仪器和试剂 兔抗人Fas、FasL多克隆抗体，为Santa Cruz公司产品。FITC标记的羊抗兔IgG抗体，购自华美生物公司。鼠抗人FasL中和性单克隆抗体NOK2、Annexin VFITC凋亡检测试剂盒及流式细胞仪FACSCan，均为美国BD公司产品。脂质体OligofectamineTMReagent，为美国Invitrogen公司产品。TRIzol Reagent、Revee Tracriptase、Taq DNA