细胞因子诱导的杀伤细胞对人胃癌高侵袭腹膜转移细胞株体内外杀瘤过程中多种细胞因子的影响  

Effects of manifold cytokine at OCUM-2MD3 cell line in vitro and vivo at cytokine-induced killer cell from healthy donors

作　　者：陈少轩[1] 赵群[2] 李勇[2] 杨进强[2] 单保恩[3] 庞志诚[4] 焦琨[3] 周宏伟[3] 

机构地区：[1]中日友好医院内镜中心微创外科组,北京100029 [2]河北医科大学第四医院外科,河北石家庄050011 [3]河北医科大学科研中心,河北石家庄050011 [4]河北医科大学第四医院药剂科,河北石家庄050011

出　　处：《中日友好医院学报》2010年第1期29-32,共4页Journal of China-Japan Friendship Hospital

基　　金：河北省中医药管理局科技攻关课题(200121)

摘　　要：目的:观察细胞因子诱导的杀伤细胞(CIK细胞)自分泌细胞因子及其体内外杀瘤过程中多种细胞因子含量的变化,探讨CIK细胞对抗肿瘤的作用机制。方法:取正常人的外周肝素抗凝血分离外周血单个核细胞(PBMC),体外诱导成CIK细胞;同时建立人胃癌高侵袭转移瘤(OCUM-2MD3)细胞裸鼠腹膜移植模型。检测CIK细胞中白介素-2(IL-2)、肿瘤坏死因子α(TNF-α)、干扰素γ(IFN-γ)、粒细胞-巨噬细胞集落刺激因子(GM-CSF)等细胞因子的分泌及其对OCUM-2MD3细胞株体内外杀瘤时上述细胞因子的变化。结果:①CIK细胞培养过程中其上清液IL-2、TNF-α、INF-γ、GM-CSF含量随时间增长而升高,至2周左右达最高峰,随后下降;②体外培养CIK细胞以E﹕T=25﹕1对OCUM-2MD3肿瘤细胞作用后,IL-2含量减低明显(P<0.05),TNF-α、GM-CSF含量均降低,INF-γ含量升高;③体内抗瘤实验结果表明,CIK治疗组血清中IL-2、TNF-α、IFN-γ、GM-CSF的含量均较生理盐水对照组显著增高(P<0.05)。结论:CIK细胞分泌的细胞因子在2周左右具有最大生物学活性;CIK细胞可能通过其自身分泌IL-2、TNF-α、IFN-γ、GM-CSF等细胞因子参与抗肿瘤作用。Objective：To investigate changes of four main eytokines during anti-tumor process of human cy-tokine-induced killer（CIK）cell in OCUM-2MD3 cell line in vitro and vivo and to discuss the possible mechanisms of CIK cell against gastric cancer.Methods：Human peripheral blood monouclear cells （PBMC） from healthy donors were separated and induced CIK cells.After the animal model was constructed,changes of cytokines were detected by radioimmuno-detection （RAID） and enzyme-linked-immunosorbent assay（ELISA）,cytotoxicity of CIK cells were analyzed in vitro by MTr assays.Results：（1）The amount of interferon-γ（IFN-γ）, interleukin-2（IL-2）,tumor necrosis factor-α（TNF-α）,granulocyte-macrophage colony-stimulating factor（GM-CSF ）were increased by the culturing time and the highest concentration appeared in the time of 2 weeks.（1） After treatment in vitro IL-2 decreased significantly（P〈0.05） whereas IFN-γ,TNF-α, GM-CSF were increased. （2） In vivo CIK cells had a stronger suppressive effect on the tumor growth in Balb/c nude mice bearing OCUM-2MD3 tumor and the concentrations of these cytokines were increase significantly compared with NS group （P〈0.05）.Conclusion：CIK cells had the most significant suppression cytotoxicity in the culturing time of 2 weeks and IFN-γ,IL-2,TNF-α,GM-CSF were correlated with the anti-tumor activity of CIK cell line in vitro and in vivo.

关 键 词：胃肿瘤 细胞因子诱导的杀伤细胞 细胞因子 人胃癌高侵袭腹膜转移细胞株 细胞毒活性 抗肿瘤 

分 类 号：R735.2[医药卫生—肿瘤]