IL-2基因转染的CIK细胞联合DC对肝癌细胞的杀伤作用  

The cytolytic effect against liver cancer cells of hIL-2 gene transfected CIK combined with DC

作　　者：匡志鹏[1] 吴继宁[1] 罗小玲[1] 谢裕安[1] 李安娜[1] 

机构地区：[1]广西肿瘤防治研究所实验研究部,南宁530021

出　　处：《中国癌症防治杂志》2009年第1期10-13,共4页CHINESE JOURNAL OF ONCOLOGY PREVENTION AND TREATMENT

基　　金：广西自然科学基金资助课题(桂科自0640158)

摘　　要：目的探讨IL-2基因转染的细胞因子诱导的杀伤细胞(CIK)联合树突状细胞(DC)对肝癌细胞的杀伤作用。方法采用密度梯度离心法从健康人外周血中分离单个核细胞(PMBC),分别诱导CIK细胞和DC。用携带IL-2基因的重组腺病毒AdBM5-GFP-IL-2转染CIK细胞,HepG2肝癌冻融抗原冲击致敏DC,然后将转染IL-2基因的CIK细胞(IL2-CIK)与肝癌抗原致敏DC(Lysate-DC)共培养,制备肝癌抗原致敏DC激活的IL-2基因修饰的CIK细胞(Lysate-DC-IL-2-CIK)。采用MTT法检测单纯CIK、DC联合CIK(DC-CIK)、肿瘤抗原致敏DC激活的CIK(Lysate-DC-CIK)、DC激活的IL2基因转染的CIK(DC-IL2-CIK)和肿瘤抗原致敏DC激活的IL-2基因转染的CIK(Lysate-DC-IL2-CIK)对肝癌HepG2细胞的杀伤作用,效靶比分别为10:1、20:1、40:1,同时以肝癌细胞H7404、白血病细胞K562作为靶细胞进行杀伤对照实验。结果研究结果表明,不同效靶比的Lysate-DC-IL-2-CIK效应细胞对HepG2的杀伤活性均显著高于对照组(P<0.05或0.01),且其杀瘤作用随着效靶比的增高而增强,在效靶比为40:1时达到最佳抗肿瘤活性。结论经肝癌抗原致敏DC激活的IL-2基因转染的CIK细胞对AFP分泌功能的肝癌细胞具有更强的杀伤作用。Objective To investigate the cytolytic effect against liver cancer cells of hIL-2 gene transfected cytokine induced killer cells （CIK） combined with dendritic cells（DC）. Methods Peripheral blood mononuelear cells （PBMC） were isolated from healthy adult donors by density gradient centrifugation. After 2 hours incubation, CIK was generated from non-adherent cells by anti-CD3McAb, IL-2, IL-1 and IFN-γ. DC was induced from adherent cells by rhIL-4 and rhGM-CSF. Recombinant adenovirus vector pAdBMS-GFP-IL-2 infected CIK at 100 MOI generated IL-2-CIK DC were pulsed with HepG2 lysate at the ration of 1 ： 3 and generate lysate-DC. The effective cells were lysate-DC-IL2-CIK, lysate-DC-CIK, CIK, DC-CIK and IL2-CIK respectively The target cells were HepG2, K562 and H7404 tumor cell line. The ratio of effective cells and target cells （E/T） were 10 ： 1, 20 ： 1 and 40 ： 1 respectively. The cytotoxicity against HepG2, K562 and H7404 was detected with MTT assays. Results There were stronger cytolytic effects against HepG2 for effective cells （Lysate-DC-IL2-CIK） than those effective cells against K562, H7404 and HepG2 in the control group at different E/T ratios. Its cytolytic effects enhanced with the rising of E/T ratio and reached climax when the E/T ratio was 1 ： 40. Conclusions hIL-2 transfected CIK combined with hepatocellular carcinoma antigen pulsed DC has more effective cytolytic effect.

关 键 词：CIK IL-2 DC 肝细胞癌 抗肿瘤作用 

分 类 号：R730.3[医药卫生—肿瘤]