摘要
目的观察细胞因子诱导的杀伤(cytokine induced killer,CIK)细胞对Lewis肺癌细胞增殖和Lewis肺癌移植瘤生长的抑制作用,探讨其抑瘤机制。方法常规方法培养CIK细胞,以CIK细胞为效应细胞,Lewis肺癌细胞为靶细胞,以不同效靶比共同培养,MTT法检测细胞增殖情况,同时电镜观察Lewis肺癌细胞形态特征改变;流式细胞仪检测Lewis肺癌细胞的凋亡;免疫细胞化学法检测并比较FasL在单个核细胞和CIK细胞表面的表达;用MTT法检测抗FasL单克隆抗体对CIK细胞杀伤Lewis肺癌细胞的影响;用ELISA方法检测Lewis肺癌荷瘤小鼠脾细胞上清液中IL-2、IL-4、IFN-γ和TNF-α浓度。结果电镜观察:CIK细胞能促进Lewis肺癌细胞凋亡超微结构的改变;流式细胞仪检测:CIK细胞组凋亡率明显高于对照组;FasL在CIK细胞表达增加;抗Fas单抗可抑制CIK杀伤Lewis肺癌细胞的活性;CIK细胞治疗的Lewis肺癌小鼠脾细胞培养上清液中细胞因子水平明显高于对照组。结论CIK细胞可在体内及体外抑制Lewis肺癌细胞增殖,诱导肿瘤细胞凋亡,Fas/FasL途径在其中发挥一定作用,同时CIK细胞抗肿瘤作用可能与淋巴细胞活化和分泌细胞因子有关。
[Objiective] To investigate the inhibitory effects of CIK on the proliferation of Lewis lung carcinoma cell line and the growth of transplanted Lewis lung carcinoma in C57BL/6N mice in vivo. [Methods] CIK ceils were induced by culturing PBMC with regular method. The proliferation of Lewis lung carcinoma ceils was measured by MTT assay. Ultramicrostructure of Lewis lung carcinoma ceils was observed under a transmission electron micro- scope. Flowcytometric analysis was used to detect cell apoptosis, uhrastructural observation expression of FasL were individually determined by MTY and immunocytochemistry (/CC) analysis. IL-2, IL-4, IFN-r and TNF-α were tested by ELISA. [Results] Electron microscopic observations showed that CIK ceils could induce the lung carcinoma ceils to apoptosis. Flow cytometrlc analysis demonstrated that apoptosis ceils of Lewis lung carcinoma increased in CIK group compared with those in the control group. FasL expression on CIK increased. Cytotoxicity was blocked after addition of anti-FasmAb. Level of cytokine in CIK therapy group was higher than that of the control group. [ Conclusions] CIK has inhibitory effect on Lewis lung carcinoma ceils both in vitro and in vivo. CIK ceils can induce the apoptosis of Lewis lung carcinoma ceils, and Fas/FasL pathway plays an important role in apoptosis of Lewis lung carcinoma cells by CIK. The CIK therapy is possibly correlated with induction of the activation of the splenic lymphocytes secreting cytokine.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2007年第21期2605-2608,共4页
China Journal of Modern Medicine
基金
黑龙江省卫生厅基金项目(No:2005-335)
黑龙江省教育厅基金项目(No:11513116)