【作者】 远洋; 王雪峰; 张扬; 王家玉; 栾宁;
【Author】 YUAN Yang1 WANG Xuefeng2 ZHANG Yang3 WANG Jiayu4 LUAN Ning2(1Department of Otolaryngology,Affiliated Hospital of Weifang Medical University,Weifang,261031,China;2Department of Otolaryngology,the First Affiliated Hospital of Liaoning Medical University;3Department of the General Surgery,Peking University Third Hospital;4Department of Otolaryngology,the First Hospital of Harbin)
【机构】 潍坊医学院附属医院耳鼻咽喉科; 辽宁医学院附属第一医院耳鼻咽喉科; 北医三院普外科; 哈尔滨市第一医院耳鼻咽喉科;
【摘要】 目的:研究SOCS1沉默的树突状细胞(DC)疫苗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DC的临床应用提供新思路。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(CTL)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时其杀伤活性与对照组之间的差异有统计学意义(P<... 更多还原
【Abstract】 Objective:Investigate the specific antitumor mechanism of SOCS1 silent DC vaccine and discuss the prospect of RNAi in the gene therapy for laryngocarcinoma in order to provide novel ideas of DC’s clinical applications.Method:Dendritic cells derived from peripheral blood monocytes were cultured in vitro in the presence of GM-CSF,IL-4 and TNF-α.The morphological feature of DC was observed with inverted microscope.RNAi vector were transfected into DC.The expression of SOCS1 protein was detected wit... 更多还原 【关键词】 细胞因子信号转导抑制因子1; RNA干扰; DC疫苗; 喉肿瘤; 【Key words】 suppressor of cytokine signaling 1; RNA interference; DC vaccine; laryngeal neoplasms; 【基金】 辽宁省自然基金项目(No:20082180) 【分类号】R739.65
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