负载抗原DC联合CIK对肝癌免疫微环境的影响  

Effect of Tumor Antigen Loaded DC-CIK on Microenvironment of Hepatocellular Carcinoma

作　　者：田大广[1] 于恒海[1] 魏晓平[1] TIAN Da-guang;YU Heng-hai;WEI Xiao-ping(The First Department of HBP Surgery,The 2nd Affiliated Hospital of Kunming Medical University,Kunming Yunnan 650101,China)

机构地区：[1]昆明医科大学第二附属医院肝胆胰外科一病区,云南昆明650101

出　　处：《昆明医科大学学报》2021年第1期46-50,共5页Journal of Kunming Medical University

基　　金：云南省教育厅科学研究基金资助项目(2019J1264)。

摘　　要：目的探讨负载肝癌抗原树突状细胞(dendritic cell,DC)联合细胞因子诱导杀伤细胞(cytokineinduced killer,CIK)对肝细胞性肝癌(hepatocellular carcinoma,HCC)微环境的影响。方法采用手术切除HCC组织制备HCC抗原,采集健康志愿者单个核细胞,诱导并分离出DC,CIK,按是否加入HCC抗原及共培养将细胞分为DC、Ag-Dc、CIK、Ag-DC-CIK四组,流式细胞仪测定各组细胞表型后,ELISA检测IL-6、IL-10及IFN-γ水平;免疫细胞化学SP法检测血管内皮生长因子(vascular endothelial growth factor,VEGF)的表达;RTPCR检测基质金属蛋白酶(matrix metalloproteinase,MMP)-2,MMP-3,MMP-9mRNA表达水平。结果负载抗原后DC组CD80及CD86表达阳性率,Ag-DC-CIK组CD3+CD56+、CD8+CD56+双阳性细胞均显著升高(P <0.05);Ag-DC-CIK组IL-6及IFN-γ浓度显著低于其余各组;而IL-10浓度最高(P <0.05);Ag-DC-CIK组VEGF及MMP-2、MMP-3、MMP-9mRNA表达水平均显著下降(P <0.05)。结论在DC负载自身抗原的基础上,与CIK共培养,可显著增强DC及CIK活性,有效降低HePG2微环境中血管形成,炎症反应及纤维化相关因子表达。Objective To study the effect of tumor antigen loaded DC-CIK on the microenvironment of hepatocellular carcinoma.Methods The antigen of HCC coming from resected HCC and peripheral blood mononuclear cells were isolated from healthy donors.The cells were divided into 4 groups as DC,Ag-DC,CIK and Ag-DC-CIK according to the loading of HCC antigen or coculture of DC-CIK or not.Cell phenotype of 4 groups were evaluated by FCM.Furthermore,levels of IL-6,IL-10 and IFN-γ;expression of VEGF and MMP-2、MMP-3、MMP-9 mRNA were observed.Results The contents of CD80,CD86 in group Ag-DC and CD3+CD56+,CD8+CD56+double positive cells in group Ag-DC-CIK elevated significantly(P<0.05).The level of IL-6 and IFN-γdecreased significantly in group Ag-DC-CIK(P<0.05).Expression of VEGF and MMP-2,MMP-3,MMP-9 mRNA also reduced significantly in group Ag-DC-CIK(P<0.05).Conclusion Coculture of DC and CIK on the basis of loading HCC antigen shows the stronger activity and reduces the expression of relating factors about inflammation,angiopoiesis and fibrosis in HePG2 microenvironment.

关 键 词：肝细胞肝癌 微环境 树突状细胞 细胞因子诱导杀伤细胞 

分 类 号：R730.54[医药卫生—肿瘤] R320.2445[医药卫生—临床医学]