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中国癌症防治杂志

中国癌症防治杂志

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IL-2基因转染的CIK细胞联合DC对肝癌细胞的杀伤作用

匡志鹏;吴继宁;罗小玲;谢裕安;李安娜;   

  1. 广西肿瘤防治研究所实验研究部;
  • 收稿日期:2009-03-25 修回日期:2009-03-25 出版日期:2009-03-25 发布日期:2009-03-25

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

KUANG Zhi-peng,WU Ji-ning,LUO Xiao-ling,et al.   

  1. (Department of Experimental Research,the Guangxi Cancer Institute, Nanning 530021,China
  • Received:2009-03-25 Revised:2009-03-25 Online:2009-03-25 Published:2009-03-25

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被引次数

14

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。

关键词: CIK, IL-2, DC, 肝细胞癌, 抗肿瘤作用

Abstract: 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 mononuclear 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 pAdBM5-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.

Key words: CIK, DC, IL-2, Hepatocellular carcinoma, Cytolytic effect