【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。更多还原

【Abstract】 Objective:To investigate changes of four main cytokines during anti-tumor process of human cytokine-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 MTT assays.Results:①The amount of interferon-γ(IFN-γ), interleukin-2(IL-2),tumor necrosis factor-α(TNF-α),granulocyte-macrophage colony-stimulating factor(GMCSF )were increased by the culturing time and the highest concentration appeared in the time of 2 weeks.② After treatment in vitro IL-2 decreased significantly(P<0.05) whereas IFN-γ,TNF-α,GM-CSF were increased. ③ 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.更多还原