目的：探讨细胞因子诱导的杀伤细胞（cytokineinduced killer cells，CIKs）对宫颈癌CasKi和Hela细胞的体内外杀伤效应。方法：从宫颈癌患者的外周血分离单个核细胞，以改良方法（不加IFNγ，只加IL2 和antiCD3抗体）刺激扩增得到CIKs，用流式细胞仪分析CIKs表型。ELISA法检测CIKs培养上清中IFNγ、IL2 和TNFα的分泌水平。LDH释放法检测CIKs对CasKi和HeLa细胞的杀伤作用。建立荷CasKi或Hela细胞小鼠模型，以1×106或1×107个CIKs小鼠静脉注射治疗，每周1次，连续治疗3周，检测小鼠移植瘤的体积和重量。结果：在抗CD3抗体和IL2的刺激下，宫颈癌患者外周血单个核细胞被诱导培养成CIKs，其中CD3+CD56+细胞得到大量扩增。PHA激活后，CIKs产生大量的IFNγ和TNFα，而IL2的分泌量较少。被激活的CIKs可显著地杀伤宫颈癌CasKi细胞和HeLa细胞，最大杀伤率分别达（43.2±1.8）%和（46.2±15）%，且呈剂量依赖性。体内抗肿瘤实验结果显示，CIKs能显著抑制小鼠皮下宫颈癌移植瘤的生长（P<0.01）。结论：本实验的改良方法能有效制备CIKs，该CIKs在体内外对宫颈癌CasKi 和 Hela细胞有明显的杀伤效应。

Objective:To investigate the antitumor activity of cytokineinduced killer cells (CIKs) against cervical cancer cell lines, CasKi and HeLa in vitro and in vivo. Methods: The CIKs were induced from peripheral blood mononuclear cells (PBMCs) of patients with cervical cancer using an improved method, which only used IL2 and antiCD3 antibody, without IFNγ; CIKs were sorted using FACS. The levels of IFNγ, IL2 and TNFα in culture supernatants of CIKs were determined by ELISA. The antitumor activities of CIKs against CasKi and HeLa cells were determined by LDH assay. The nude mouse xenograft models of cervical cancer cell lines, CasKi or HeLa cells, were established, and 1×106 or 1×107 CIKs were administered intravenously once a week for three weeks, then the tumor volumes and weights were measured.Results: CIKs were successfully induced from PBMCs of cervical cancer patients by IL2 and antiCD3 antibody, with CD3+CD56+ cells greatly expanded. CIKs produced significant amounts of IFNγ and TNFα, but few IL2, after PHA stimulation. CIKs dosedependently killed CasKi and Hela cells with a maximum killing rate reaching 43% and 46%, respectively. In addition, the in vivo antitumor experiments demonstrated that CIKs remarkably inhibited the growth of subcutaneous tumors in nude mice (P<0.01). Conclusion: The improved CIKs expansion method used in our study is feasible and the resultant CIKs have remarkable cytotoxicity against Caski and HeLa cells both in vivo and in vitro.

武汉市创新人才开发基金资助项目(武人社函\[2009\]97号）