负载人结肠癌Lovo细胞总RNA抗原树突状细胞对细胞因子诱导的杀伤细胞在体外特异性杀伤的影响  

Effect of dendritic cells transfected with total RNA of human colon cancer Lovo cells on specific cytotoxicity of cytokine-induced killer cells in vitro

作　　者：刘季芳[1] 祁岩超[1] 杨波[1] 卢敏莹[1] 潘东晓[1] 申鸿卓[1] 

机构地区：[1]广州医学院附属肿瘤医院实验中心,510095

出　　处：《国际肿瘤学杂志》2010年第6期477-480,共4页Journal of International Oncology

基　　金：基金项目：广州医学院博士启动基金（0706078）

摘　　要：目的探讨人结肠癌Lovo细胞总RNA抗原致敏的树突状细胞（DC）对细胞因子诱导的杀伤细胞（CIK）在体外特异性杀伤的影响。方法利用Ficoll密度梯度离心法提取脐血单核细胞，分别诱导CIK和DC细胞，并用流式细胞仪检测其免疫表型。采用Trizol提取结肠癌Lovo细胞总RNA作为肿瘤细胞抗原，转染脐血来源的DC。实验分为3组：转染LovoDC共培养CIK组、未转染DC共培养CIK组和单纯CIK组。靶细胞为Lovo细胞，在效靶比为50：1和20：1的条件下以噻唑蓝法分别检测CIK的体外杀伤活性。结果在效靶比为20：1时，负载LovoRNA抗原的DC能诱导出CIK对Lovo细胞最强的细胞毒杀伤力为（76．49±4．21）％，DC＋CIK组次之为（53．84±2．15）％，CIK组细胞毒性最低为（32．20±3．07）％，且两组间差异有统计学意义（P〈0．05）。结论肿瘤细胞总RNA提取方法简单，易于临床实施，其作为抗原致敏DC能强化CIK的特异性杀伤，将有很好的I临床应用前景。Objective To explore the effect of dendritic cells （DC） primed by total RNA extracted from human colon cancer Lovo cell on specific cytotoxicity of eytokine-indueed killer cells （CIK） in vitro. Methods Cord blood mononuelear cells extracted by Ficoll density gradient centrifuge were induced into CIK and DC cells separately, and their Immunophenotype was detected by Flow eytometer. Trizol harvested total RNA from colon cancer cell Lovo and the RNAs were loaded to DCs obtained from cord blood as tumor anti gens. Effectors were grouped accordingly as CIK cells co-cultured with DCs transfected with Lovo RNA, CIK cells cocultured with unloaded DCs and CIK cells. Targets was Lovo cells. In vitro cytotoxicity of CIK cells was examined by MTY assay when the ratio of effectors to targets was 50 ： 1 and 20 ： 1 seperately. Results When the ratio of effectors to targets was 20 ： 1, the strongest cytotoxicity against Lovo ceils was achieved by CIKs cocultured with DCs loaded with Lovo RNA（76.49%±4.21% ）, DC ＋ CIK group was lower（53.84%±2.15% ）, and CIK cells group possessed the lowest cytotoxicity（32.20%±3.07% ）, showing statistic significance （ P 〈 0.05 ）. Conclusion Extraction of total RNA from tumor cells is simple and easy for clinical implementation. Total RNAs acted as antigen to pulse DCs can strengthen the specific cytotoxicity of CIK cells, which will have good prospects for clinical application.

关 键 词：树突细胞 杀伤细胞 抗原 

分 类 号：R735.35[医药卫生—肿瘤]