FasL及FasLASODN在舌鳞癌细胞免疫逃逸中的作用

摘要

Objective To research the effects of FasL and FasL ASODN in the immune escape of squamous lingual carcinoma cells .Methods This experiment was planned to conduct the test through the expression and functions of FasL in squamous lingual carcinoma cells ,and designed and compounded the specific FasL ASODN to transfect squamous lingual carcinoma cells and block the expression of FasL in squamous lingual carcinoma cells .Such methods as RT‐PCR ,flow cytometry and M T T had been adopted to conduct the observation from three levels ,gene ,protein and cell effects .Results The FasL ASO inhibited cell proliferation in a dose‐and time‐dependent manner within limits(P<0 .05) .The total apoptosis rate was apparently increased(P<0 .05) in antisense group compared with the control group ,the lipofectamine group and the sense group .Conclusion Fas/FasL approach is one of the important mechanisms of the immune escape of oral squamous cell carcinoma cells ,and can be the new target for immunotherapy .%目的：研究Fas配体（FasL）及FasL反义寡脱氧核苷酸（ASODN）在舌鳞癌细胞免疫逃逸中的作用。方法对人舌鳞癌细胞中FasL表达及功能进行检测，设计合成特异FasLASODN，转染舌鳞癌细胞，封闭舌鳞癌细胞的FasL表达。通过流式细胞术、逆转录聚合酶链反应（RT‐PCR）和四唑盐（MTT）等方法，观察基因‐蛋白‐细胞效应。结果通过RT‐PCR检测提示在脂质体的量一定的情况下，在一定范围内，ASO的浓度越高，转染效率越高。MTT／FCM结果显示ASO对SCC‐9细胞的增殖有明显的抑制作用（P＜0．05）。FCM对细胞凋亡的检测结果显示反义链组与正义链组、空白对照组及脂质体组相比，总凋亡率有明显差异（P＜0．05）。结论Fas／FasL途径是口腔鳞癌细胞免疫逃逸的重要机制之一，可作为免疫治疗的新靶点。