CIK细胞体内外抗宫颈癌HeLa细胞活性的研究

目的：探讨细胞因子诱导的杀伤（cytokine-induced killer,CIK）细胞的体内外抗宫颈癌HeLa细胞活性。方法：收集8名健康献血者和8名宫颈癌患者的新鲜外周血,分别通过常规方法分离外周血单核细胞（peripheral blood mononuclear cells,PBMC）,应用相应细胞因子体外诱导分化出CIK细胞,动态观察CIK细胞的体外增殖活性、细胞表型和对HeLa细胞的杀伤活性;在BALB/c裸鼠皮下接种效应细胞,观察宫颈癌患者CIK细胞对接种HeLa细胞的荷瘤鼠的抑瘤作用,同时设淋巴因子激活的杀伤细胞（lymphokine activated killer cells,LAK）和PBMC细胞作为对照。结果：源于健康人和宫颈癌患者的CIK细胞间的增殖活性无明显区别（P〉0.05）。表型分析结果表明,两种来源的CIK细胞中CD3~＋CD56~＋双阳性细胞均得到了大量扩增,宫颈癌患者CIK细胞中CD3~＋CD56~＋双阳性细胞在实验开始前约占0.13%,到实验后第28天上升到25.8%。体外实验表明,宫颈癌患者的CIK细胞杀伤宫颈癌HeLa细胞的细胞毒活性明显高于PBMC细胞。裸鼠体内实验表明,宫颈癌患者CIK细胞能够显著抑制肿瘤的生长,其抑瘤率可达80.6%,高于LAK细胞的59.1%和PBMC细胞的38.3%（P〈0.01）。CIK治疗后肿瘤体积明显比空白对照组缩小（P〈0.05）。结论：宫颈癌患者CIK细胞具有较强的体内外抗宫颈癌细胞活性,有可能用于临床上宫颈癌的过继性免疫治疗。

Antitumor effects of cytokine- induced killer cells on cervical cancer HeLa cells in vitro and in vivo

OBJECTIVE:To explore the differences of cytokine-induced killer(CIK) cells from healthy persons and patients with primary cervical cancer,in their phenotype,proliferation activity,cytotoxic activity and antitumor effects in vitro and in vivo.METHODS:CIK cells were generated by IFN-γ,CD3McAb,IL-2,IL-1 induction of cultured peripheral blood mononuclear cells(PBMC) of both 8 healthy blood donors and 8 cervical cancer patients.These treated cells were followed at different intervals by MTT assays and flow cytometry analysis.The antitumor activity of the CIK,LAK and PBMC cells were evaluated in BALB/c nude mice bearing HeLa cervical cancer. RESULTS:There was no significant difference in proliferation performance of CIK cells between healthy persons and cervical cancer patients(P>0.05).The flow cytometry analysis showed that the percentage of CD3~+CD56~+ cells increased from 0.13%on day 0 to 25.8%on day 28.CIK cells generated from patients with cervical cancer patients possessed a higher antitumor cytotoxic activity in vitro than PBMC.In addition,CIK cells had a stronger suppressive effect on tumor growth in BALB/c nude mice bearing cervical cancer than LAK cells(median inhibitory rates 80.6%vs 59.1%, respectively,P<0.01) or PBMC(median inhibitory rates 80.6%vs 38.3%,respectively,P<0.01).The tumor size in the experiment group was smaller than that in the control group after CIK treatment(P<0.05).CONCLUSION:CIK cells had a significantly stronger suppression on growth of cervical cancer cells,providing an experimental basis for CIK clinical use as an adoptive immunotherapy.