干扰Foxp3在神经母细胞瘤细胞SK-N-BE2中的表达及对其免疫功能的影响  

Effects of knocking down Foxp3 on immunological functions of human neuroblastoma SK-N-BE2 cell

作　　者：孙婧[1] 唐锁勤[2] 

机构地区：[1]解放军总医院第一附属医院儿科,北京100037 [2]解放军总医院儿科

出　　处：《中华实用儿科临床杂志》2013年第3期175-177,共3页Chinese Journal of Applied Clinical Pediatrics

基　　金：国家高技术研究发展计划（863计划）（2012AA020804）

摘　　要：目的探讨降低Foxp3的表达对神经母细胞瘤细胞SK—N—BE2免疫功能的影响。方法设计、合成Foxp3的siRNA干扰序列，转染神经母细胞瘤细胞SK—N—BE2，分别采用ReM—timePCR及流式细胞术观察Foxp3以及共刺激分子CD86在干扰前后的表达；最后将Foxp3干扰的SK—N—BE2细胞与人外周血共培养，观察Foxp3的不同表达水平对上述培养体系中T淋巴细胞功能的影响。结果本研究设计、合成的siRNA显著降低了Foxp3在神经母细胞瘤细胞SK—N-BE2中的表达，低表达Foxp3的SK—N—BE2细胞上共刺激分子CD86的表达显著升高，且在共培养体系中细胞因子IFN-γ、IL-17的表达出现轻微上调，但同时显著降低了免疫抑制分子IL-10、TGF—β在mRNA水平的表达。结论Foxp3在神经母细胞瘤中的高表达可能是该细胞逃避免疫监视的重要机制，干扰Foxp3的表达能降低共培养体系中的免疫抑制分子表达，表明干预该分子可望为肿瘤的免疫防治提供新的思路。Objective To investigate whether knocking down Foxp3 affects the immunological function of neuroblastoma cells. Methods Different siRNAs of Foxp3 were designed and the one that effectively inhibited Foxp3 gene expression was selected by real-time polymerase chain reaction and by fluorescence activated cell sorter analysis. Effects of Foxp3 knocking down on the expression of co-stimulatory molecule CD86 were also examined. Finally, neuroblastoma cells silenced of Foxp3 expression were co-cultured with human peripheral blood monouclear cells and the activities of T cells were examined. Results Expression of Foxp3 in SK-N-BE2 cells could be efficiently knocking down in both mRNA and protein levels. Knocking down Foxp3 increased the expression of CD86 and moderately increased IFN-γ and IL- l7 expressions, and significantly inhibited the expressions of TGF-β and IL-10. Conclusion Silencing Foxp3 expression in neuroblastoma SK-N-BE2 cells reverses its immune evasion mechanisms ,which suggests that this is a new strategy for treating this kind of diseases.

关 键 词：FOXP3 小干扰RNA 神经母细胞瘤 SK—N—BE2细胞株 

分 类 号：R739.4[医药卫生—肿瘤]