SOCS1沉默增强树突状细胞抗喉癌免疫效应  

Silencing of SOCS1 enhances DC-mediated anti-laryngocarcinoma immunity

作　　者：远洋[1] 王雪峰[1] 张扬[2] 江祺川[1] 李兵[3] 

机构地区：[1]辽宁医学院第一附属医院耳鼻喉科,锦州121001 [2]北京大学第三医院普外科,100191 [3]辽宁医学院附属第一医院眼科,锦州121001

出　　处：《免疫学杂志》2011年第9期794-798,共5页Immunological Journal

基　　金：辽宁省自然基金(20082180)

摘　　要：目的研究SOCS1沉默的树突状细胞特异性抗肿瘤作用机制,并探讨RNAi技术在喉癌基因治疗中的应用前景,为树突状细胞的临床应用提供新思路和理论依据。方法以细胞因子GM-CSF、IL-4和TNF-α体外诱导扩增外周血单核细胞来源的DC,倒置显微镜下观察DC形态特征;构建RNAi载体转染DC,Western blot检测SOCS1的表达情况,筛选抑制SOCS1表达的有效靶序列;流式细胞术检测DC表面分子CD83、CD86和HLA-DR的表达;ELISA法分析上清中IFN-γ的含量;MTT法评估DC刺激T细胞增殖的能力及诱导细胞毒性T细胞的杀伤活性。结果 DC体外诱导培养成功;设计的RNAi载体经测序验证无误。干扰序列5可显著下调SOCS1表达水平;SOCS1沉默联合喉癌Hep-2抗原致敏的DC可显著上调表面分子标志CD83(85.61±0.96)%、CD86(96.86±1.20)%和HLA-DR(98.02±0.94)%的表达;该组DC能有效刺激T细胞增殖,增加IFN-γ的分泌量,最终增强CTL的特异性杀伤作用,效靶比为50:1时其杀伤活性显著高于对照组(P<0.01)。结论 SOCS1沉默并负载喉癌Hep-2抗原的DC可以产生高效而特异性的抗喉癌免疫应答。This research aims to investigate the specific antitumor mechanism of suppressor of cytokine signaling 1（SOCS1） silent dendritic cells（DC） and to study the value of RNA interference technique in gene therapy for laryngocarcinoma in order to provide novel ideas and basis of DC＇s clinical applications.First,DC derived from peripheral blood monocytes were cultured in vitro at the presence of GM-CSF,IL-4 and TNF-α,then observed with inverted microscope.Hep-2 antigen was successful obtained via high-speed centrifugation after repeated freezing and thawing.The results of flow cytometry showed SOCS1 silent DC which were loaded with Hep-2 antigen had high expressions of CD83（85.61%±0.96%）,CD86（96.86%±1.20%） and HLA-DR（98.02%±0.94%）.SOCS1 silent DC could also stimulate the proliferation of T cells effectively as well as increase the production of IFN-γ,eventually enhance the specific killing effect of CTL.The enhanced killing activity was significantly higher than that of control group when the effect cells and target cells were combined at the ratio of 50：1（P 0.01）.To sum up,SOCS1 silent DC loaded with Hep-2 antigen could induce effective and specific anti-laryngocarcinoma immune responses.

关 键 词：细胞因子信号转导抑制因子1 小干扰RNA 树突状细胞 喉癌 

分 类 号：R739.65[医药卫生—肿瘤]