DC-CIK细胞对人肝癌HEPG2细胞周期,凋亡的影响.docVIP

PAGE 2 PAGE 1 基金项目：重庆市卫生局医学科学技术研究项目(2010-2-301) 作者简介：项颖，男，重庆市人，主任医师。主要从事肿瘤发病机制及其治疗方面的研究。E-mail: xiang519@ 通信作者：秦波，电话E-mail：cqqinbo@126.com DC-CIK细胞对人肝癌HEPG2细胞周期、凋亡的影响 项颖 秦波 田小利(重庆医科大学附属第一医院感染科 重庆400030) 摘要 目的 探讨细胞因子从肝癌患者外周血贴壁单个核细胞(peripheral blood mononuclear cells,PBMCs) 诱导的树突状细胞（dendritic cells，DCs）与杀伤（cytokine induced killer,CIK）细胞对人肝癌HEPG2细胞系周期、凋亡的影响。方法 用肝癌患者肝癌组织裂解物（肿瘤抗原）致敏经GM-CSF、TNF-ａ及IL-4等诱导该患者PBMC产生的DCs。用IFN-γ、IL-2和人CD3 单克隆抗体（human CD3 monoclonal antibody,hCD3mAb)等诱导该PBMC的悬浮细胞产生CIK细胞。用Tanswell培养小室将DC-CIK细胞与HEPG2细胞在同一培养体系内经该小室膜分隔培养24、48 h。流式细胞仪检测该HEPG2细胞的周期；Annexin V-PI双染法检测其凋亡。RT-PCR、Western blot分别检测凋亡相关基因Bax及其编码蛋白的表达。结果 DC-CIK细胞可明显影响人HEPG2细胞生长周期，两者共培养，可将HEPG2细胞系阻滞于G1期。DC-CIK细胞能显著诱导HEPG2细胞的凋亡(与对照相比，其凋亡诱导率差异显著，P0.01)。基因与蛋白水平检测表明，DC-CIK细胞可致HEPG2细胞系的凋亡相关基因Bax表达明显上调。结论 肝癌患者DC-CIK细胞可明显抑制肝癌HEPG2细胞系的生长，显著诱导该细胞凋亡，且该凋亡诱导具“时间-依赖”与“细胞数量-依赖”特性。 关键词 DC-CIK细胞 肝癌HEPG2细胞 细胞周期 细胞凋亡 The effect of CIK cells combined with dendritic cells of patient with liver cancer on the cycle and apoptosis of human HEPG2 cell line XIANG Ying QIN Bo TIAN Xiaoli(Department of Infectious Diseases,The First Affiliated Hospital of Chongqing Medical University, Chongqing 400030, China ) Abstract: Objective To explorate the effect of cytokine induced killer (CIK）cells and dendritic cells（DCs）from peripheral blood mononuclear cell (PBMC) of patient with liver cancer on the cycle and apoptosis of human HEPG2 cell line of liver cancer. Methods The adherent PBMCs were induced by GM-CSF、TNF-ａand IL-4 in order to produce DCs, which were sensitized with antigen of autologous tissue of liver cancer. Meanwhile, CIK cells were obtained from the suspended PBMCs induced by IFN-γ、IL-2 and human CD3 monoclonal antibody（hCD3mAb). The DCs, CIK cells and HEPG2 cell line were cultured in a same circumstance with a culturing cabin named Transwell，but the DCs-CIK cells were cultured at one side of the membrane of the Transwell, HEPG2 cell line at another side of the same membrane for 24 h and