Cholangiocarcinoma-derived exosomes inhibit the antitumor activity of cytokine-induced killer cells by down-regulating the secretion of tumor necrosis factor-α and perforin  

胆管癌外泌体通过抑制肿瘤坏死因子α与穿孔素分泌使细胞因子诱导的杀伤细胞抗肿瘤活性下降（英文）

作　　者：Jiong-huang CHEN Jian-yang XIANG Guo-ping DING Li-ping CAO 

机构地区：[1]Department of General Surgery, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University

出　　处：《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》2016年第7期537-544,共8页浙江大学学报（英文版）B辑（生物医学与生物技术）

基　　金：Project supported by the National Natural Science Foundation of China(No.81272671);the Foundation of Health and Family Planning Commission of Zhejiang Province(No.2015KYB218);China

摘　　要：Objective： The aim of our study is to observe the impact of cholangiocarcinoma-derived exosomes on the antitumor activities of cytokine-induced killer （CIK） cells and then demonstrate the appropriate mechanism. Methods： Tumor-derived exosomes （TEXs）, which are derived from RBE cells （human cholangiocarcinoma line）, were collected by ultracentrifugation. CIK cells induced from peripheral blood were stimulated by TEXs. Fluorescence-activated cell sorting （FACS） was performed to determine the phenotypes of TEX-CIK and N-CIK （normal CIK） cells. The concen- trations of tumor necrosis factor-a （TNF-a） and perforin in the culture medium supematant were examined by using an enzyme-linked immunosorbent assay （ELISA） kit. A CCK-8 kit was used to evaluate the cytotoxic activity of the CIK cells to the RBE cell line. Results： The concentrations of TNF-a and perforin of the group TEX-CIK were 138.61 pg/ml and 2.41 ng/ml, respectively, lower than those of the group N-CIK 194.08 pg/ml （P〈0.01） and 3.39 ng/ml （P〈0.05）. The killing rate of the group TEX-CIK was 33.35%, lower than that of the group N-CIK （47,35% （P〈0.01））. The population of CD3＋, CD8＋, NK （CD56＋）, and CD3＋CD56＋ cells decreased in the TEX-CIK group （63.2±6.8）%, （2.5±1.0）%, （0.53±0.49）%, （0.45±0.42）%） compared with the N-CIK group （（90.3±7.3）%, （65.7±3.3）%, （4.2±1.2）%, （15.2±2.7）%）, P〈0.01. Conclusions： Our results suggest that RBE cells-derived exosomes inhibit the antitumor activity of CIK cells by down-regulating the population of CD3＋, CD8＋, NK （CD56＋）, and CD3＋CD56＋ cells and the secretion of TNF-a and perforin. TEX may play an important role in cholangiocarcinoma immune escape.目的:探索胆管癌来源外泌体(TEX)对细胞因子诱导的杀伤细胞(CIK)抗肿瘤活性的影响,并初步探讨其作用机制。创新点:首次通过体外实验证明TEX可引起CIK抗肿瘤活性下降,且此作用与肿瘤坏死因子α(TNF-α)和穿孔素表达抑制相关。方法:采用超速离心法提取人胆管癌细胞(RBE)来源的外泌体,同时CIK通过人外周血培养获得。将TEX负载到CIK培养体系中作为TEX-CIK组,不加TEX的CIK作为N-CIK组。流式细胞仪检测两组CIK细胞表型变化,酶联免疫吸附法(ELISA)检测两组培养基上清液中TNF-α和穿孔素的浓度,CCK-8法检测CIK对RBE细胞的杀伤活性。结论:TEX能降低CIK细胞CD3^+、CD8^+、NK(CD56^+)以及CD3^+CD56^+比例,并且抑制TNF-α和穿孔素表达,从而降低CIK细胞的抗肿瘤效应。

关 键 词：CHOLANGIOCARCINOMA Tumor-derived exosomes Cytokine-induced killer cells Immune escape 

分 类 号：R735.8[医药卫生—肿瘤]