CIK细胞对肺腺癌细胞株A549体内外杀瘤活性的实验研究

Anti-tumor Effects of CIK Cells Against Lung Cancer Cell Lines A549 in Vitro and in Vivo

目的探讨正常人细胞因子激活的杀伤(cytokine induced-killer,CIK)细胞的表型变化、增殖活性及对人肺腺癌细胞株A549细胞的体内外杀伤作用。方法取健康人外周血单个核细胞(PBMC),经IFNγ-、IL-2、抗CD3单抗联合诱导CIK细胞,观察CIK细胞增殖活性,流式细胞仪测其细胞表型变化及四甲基偶氮唑蓝(MTT)法测定其体外的细胞毒活性,用肺腺癌细胞株A549建立裸鼠肺癌模型观察其体内的抗肿瘤效果。结果培养21 d内,CIK细胞增殖(19.7±4.2)倍;CD3+CD56+表达水平明显上调,第14天时达(26.3±3.6)%;体外实验表明CIK细胞对A549细胞有明显细胞毒活性,效靶比为20:1时杀伤率为(55.4±6.2)%。体内实验表明,CIK细胞可有效抑制裸鼠皮下肺癌移植瘤的生长(P<0.01)。结论CIK细胞是一种高效的免疫效应细胞,能够显著抑制体内外肺腺癌细胞株A549的生长,为肺癌免疫治疗提供了一种新的治疗手段。

Objective To investigate the changes of phenotypes, proliferation activity and cytotoxicity of the human cytokine induced killer（CIK） cells from healthy donor against lung cancer cell lines A549 in vitro and in vivo. Methods Peripheral blood mononuclear cells（PBMC） from healthy donors were incubated to induce CIK cells in the presence of interferon gamma（IFN-γ）, IL-2 and anti-CD3 monoclonal antibody（mAb）. The phenotypes changes of CIK cells were identified by flow cytometric analysis. MTT assays were used to determine the cytotoxicity of CIK cells. The antitumor activity of CIK cells were evalulated in BALB/c nude mice bearing A549 lung cancer. Results The total CIK cells significantly increased by （19.7 ±4.2） fold in cell proliferation number at day 21 incubation. The expression rate of CD3^＋ CD56^＋ cells rose from （4.9 ± 1.4） % to （26.3 ± 3.6） % on the 14th day. In vitro, CIK cells demonstrated （55.4 ± 6.2） % lysis of A549 at an effector： target ratio of 20 ： 1. In tumor bearing nude mice, CIK cells showed a significant inhibitory effect on the growth of transplanted lung cancer cell, resulting in a statistical significant difference compared with the control group. Conclusion CIK cells are of highly efficient cytotoxic effector cells against lung cancer cell lines A549 in vitro and in vivo and are likely to be usedas an immune therapeutic strategy for lung cancer.