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目的探讨恶性肿瘤对于细胞因子诱导的杀伤(cytokine induced killer,CIK)细胞的增殖抑制以及诱导凋亡等方面的作用,并探讨肿瘤细胞影响CIK的作用机制。方法在CIK细胞培养体系中分别加入胃癌细胞系BGC-823、结肠癌细胞系HCT-8细胞的培养上清,观察对于CIK细胞的增殖倍数、主要效应细胞(CD3~+ CD56~+细胞)的含量以及杀伤活性的影响。为探讨肿瘤细胞来源的胞外体在影响CIK细胞活性中的作用,采用超速离心技术分离肿瘤细胞培养上清中的胞外体结构,观察其对于CIK细胞的抑制作用。结果培养体系中肿瘤培养上清的加入降低了CIK细胞的增殖倍数和CD3~+ CD56~+细胞的比例,并且肿瘤培养上清的加入抑制了CIK细胞对于同类肿瘤细胞的细胞毒活性。肿瘤细胞和CIK细胞的共同培养增加了CIK细胞中凋亡细胞的比例,同时降低了增殖细胞比例。通过超速离心方法从BGC-823胃癌细胞培养上清液中分离出胞外体结构,电镜显示为直径介于40～100 nm之间的膜性微小囊泡,并且BGC-823细胞培养上清中胞外体结构的去除可以减轻其对于CIK细胞的增殖抑制。结论肿瘤细胞对于CIK细胞的增殖和生物学活性具有抑制作用,免疫抑制性细胞因子和胞外体都是可能的作用机制。
Objective To investigate the effects of malignant tumor on proliferation and apoptosis induction of cytokine induced killer (CIK) cells and to explore the mechanism of action of tumor cells on CIK. Methods The culture supernatants of gastric cancer cell line BGC-823 and colon cancer cell line HCT-8 were respectively added into CIK cell culture system. The proliferation of CIK cells and the expression of major effector cells (CD3 ~ + CD56 ~ +) Content and killing activity. To investigate the role of extracellular matrix of tumor cells in influencing the activity of CIK cells, the extracellular body structure in the culture supernatant of tumor cells was isolated by ultracentrifugation and its inhibitory effect on CIK cells was observed. Results The addition of tumor culture supernatant reduced the multiplication ratio of CIK cells and the proportion of CD3 ~ + CD56 ~ + cells, and the addition of tumor culture supernatant inhibited the cytotoxic activity of CIK cells against the same tumor cells. Coculture of tumor cells and CIK cells increased the proportion of apoptotic cells in CIK cells while decreasing the proportion of proliferating cells. The extracellular structure was isolated from BGC-823 gastric cancer cell culture supernatant by ultracentrifugation. Electron microscopy showed membranous microvesicles between 40 and 100 nm in diameter, and in the culture supernatant of BGC-823 cells Removal of extracellular structures can reduce its inhibition of proliferation of CIK cells. Conclusions Tumor cells have inhibitory effects on the proliferation and biological activity of CIK cells. Both immunosuppressive cytokines and extracellular bodies are possible mechanisms.