摘要
目的探讨细胞因子激活的杀伤细胞(cytokine-induced killer cells,CIK)对人食管癌EC9706细胞裸鼠皮下移植瘤的抑制作用。方法体外分离健康人外周血单个核细胞,干扰素-γ、白细胞介素-2、CD3单抗诱导培养14 d,流式细胞仪测细胞表型,LDH法测定不同效靶比时CIK细胞对EC9706细胞的杀伤活性。12只BALB/c裸鼠分为两组,每组6只。每只裸鼠皮下接种1×106个EC9706细胞,治疗组裸鼠同时每只经尾静脉注入3×107个CIK细胞,每周1次,连续3周,观察两组裸鼠成瘤时间、成瘤率、肿瘤体积变化。成瘤后3周,眼眶取血,流式细胞仪检测人CIK细胞;处死裸鼠,取瘤块称重,计算抑瘤率,观察肿瘤病理特点。结果CIK细胞中CD3+CD56+细胞占42.50%,CIK细胞表面NKG2D表达率为67.10%。效靶比10∶1、20∶1、30∶1时CIK细胞对EC9706细胞的杀伤活性分别为(28.81±0.47)%、(37.78±0.22)%、(44.31±1.06)%,差异有统计学意义(P<0.01)。对照组和CIK细胞治疗组成瘤时间分别为(9.00±1.26)d、(15.67±4.37)d,差异有统计学意义(P<0.01);瘤重分别为(3.24±0.11)g、(2.10±0.10)g,差异有统计学意义(P<0.01),CIK治疗组的抑瘤率为35.19%。HE染色显示CIK细胞治疗组有较多的淋巴细胞浸润和坏死区。结论CIK细胞对EC9706细胞裸鼠皮下移植瘤有明显的抑制作用,可能成为治疗食管癌的免疫效应细胞。
Objective To investigate the inhibitory effect of cytokine-induced killer（CIK）cells against the human esophagus carcinoma cells EC9706 transplanted tumor in nude mice.Methods CIK cells were generated from peripheral mononuclear cells of a healthy volunteer in the presence of IFN-γ,CD3 mAb and IL-2 in vitro.The immune phenotype of CIK cells 14 days after culture were analyzed by flow cytometry.Cytotoxicity of CIK cells against EC9706 cells were detected by LDH releasing assay at different effect-to-target cell ratios（E∶T）.12 nude mice were divided into two groups（the control group and the treatment group）,6 nude mice in the treatment group were injected subcutaneously 1×10^6 EC9706 cells and 3×10^7 CIK cells intravenously weekly,while 6 nude mice in the control group were just injected 1×10^6 EC9706 cells subcutaneously.3 weeks after the treatment,the tumor formation time,the tumor formation rate,the changes of tumor volume were observed.3 weeks after tumor formation,all the mice were euthanized,human CIK cells in peripheral blood were analyzed by flow cytometry and the tumor weights were measured.Results Of CIK cells,42.50% expressed CD3＋CD56＋,67.10% expressed NKG2D.Cytotoxicity of CIK cells against EC9706 cells were（28.81±0.47）%,（37.78±0.22）%,（44.31±1.06）%,respectively at 10∶1,20∶1,30∶1 E∶T ratios（P〈0.01）.CIK cells obviously inhibited the growth of EC9706 transplanted tumor in nude mice,the time of tumor formation in the control group and the treatment group were（9.00±1.26） d,（15.67±4.37） d,respectively（P〈0.01）.the average weight of tumors in the control group and the treatment group were（3.24±0.11） g,（2.10±0.10） g,respectively（P〈0.01）.The tumor growth inhibition rate was 35.19% in the treatment group.HE stain showed that there were more lymphocytes infiltration and necrosis regions in tumor tissues in the treatment group than that in the control group.Conclusion CIK cells can inhibit EC9706 cells transplanted tumor in nude mice,CIK cells can be immune effector cells for the treatment of esophagus carcinoma.
出处
《肿瘤基础与临床》
2010年第1期4-7,共4页
journal of basic and clinical oncology
基金
郑州市科技计划攻关项目(编号:083SGYS33238-1)
关键词
食管癌
CIK细胞
裸鼠
移植瘤
细胞治疗
esophagus carcinoma
cytokine-induced killer cells
nude mice
transplanted tumor
cell therapy
