CIK细胞体内外抗肝癌细胞作用

目的：研究肝癌患者CIK（cytokine-induced killer) 细胞的体外杀伤自体肝癌原代细胞的细胞毒活性以及正常人CIK细胞在裸鼠体内的抗肿瘤作用。方法：分别分离获得肝癌患者和下人的外周血单个核细胞（PBMC），加入细胞因子，体外诱导成CIK细胞，用流式细胞仪对细胞作动态表型分析，并与正常人的CIK细胞作对比。用^51Cr释放法，测定肝癌患者的CIK细胞体外杀伤自体肿瘤细胞的细胞毒性活性。在Balb/c裸鼠皮下接种肝癌细胞BEL－7402，观察CIK细胞对荷瘤鼠的抑瘤作用，并与LAK、PBMC细胞相对比。结果：肝癌患者的CIK细胞体外增殖力强，至培养28天时达到最大增值倍数300多，表型分析结果表明，CD^3 CD56^ 双阳性细胞得到了大量的扩增，其含量由原来的0．23％上升到第21天的17．8％。体外实验表明，肝癌患者的CIK细胞杀伤自体原代肝癌细胞的细胞毒性活性明显高于自体的PBMC细胞。裸鼠体内实验表明，肝癌患者的CIK细胞能够显著抑制肿瘤的生长，其抑瘤率可达84．7％，高于LAK细胞的52．8％及PBMC的37．1％（P＜0．05和P＜0．01）。结论：CIK细胞具有较强的体内外抗肝癌细胞活性，有可能应用于临床上肝癌的过继性免疫治疗。

Antitumor effects of CIK cells against hepatocellular carcinoma cells in vitro and in vivo

Purpose:To evaluate the antitumor effects of human cytokine-induced killer (CIK) cells from healthy donors and patients with primary hepatocellular carcinoma (HCC) in vitro and in vivo .Methods:Peripheral blood mononuclear cells (PBMC) from healthy donors and patients were induced to become CIK cells by in vitro incubation. The CIK cells were identified at different intervals by flow cytometry analysis. The cytotoxicity of CIK cells against autologous primary HCC was determined by 51Cr Release Assays. The antitumor activity of the CIK, LAK and PBMC cells were evaluated in Balb/c nude mice bearing BEL-7402liver cancer.Results:The flow cytometry analysis showed that CIK cells from patients with HCC proliferated and expanded more than 300-fold in vitro incubation by day 28;the percentage of CD3 CD56 cells increased from 0.23% on day 0 to 17.8% on day 21. CIK cells generated from patients with HCC patients possessed a higher antitumor cytotoxic activity on autologous HCC cells in vitro than autologous PBMC. In addition, CIK cells had a stronger suppressive effect on the tumor growth in Balb/c nude mice bearing BEL-7402 tumor than LAK cells (median inhibitory rates 84.7% vs 52.8%, P <0.05) or PBMC (median inhibitory rates 84.7% vs 37.1%, P <0.01). Conclusions:CIK cells have a stronger significant suppression against growth of primary and secondary HCC cells. The result provides an experimental basis for CIK clinical application as a adoptive immunotherapy.