Hyper-IL-6基因诱导的体内抗肝癌免疫功能  

Induction of anti-hepatocellular carcinoma immunity by Hyper-IL-6 gene in vivo

作　　者：郄春花[1] 李顺天[1] 曹武奎[1] 陆伟[1] 梁树人[1] 

机构地区：[1]天津市传染病医院天津市肝病医学研究所,天津300192

出　　处：《细胞与分子免疫学杂志》2012年第9期899-902,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：天津市卫生局基金项目(05ky02)

摘　　要：目的:建立稳定表达Hyper-IL-6基因的小鼠肝癌细胞株,探讨Hyper-IL-6用于诱导抗肝癌主动免疫治疗的可行性。方法:用脂质体法将Hyper-IL-6基因转染小鼠肝癌细胞系MM45T.Li,通过G418筛选获得抗性细胞株,命名为MM45T-HIL-6,RT-PCR和ELISA法检测目的基因的表达。同时筛选空载体pEGFP-C1转染的阳性克隆命名为MM45T-mock作为对照。将BALB/c小鼠随机分为3组,于各组小鼠右侧腋部皮下分别接种细胞MM45T.Li、MM45T-mock和MM45T-HIL-6 5×105个/只,体内实验观察MM45T.Li、MM45T-mock及MM45T-HIL-6的成瘤性,流式细胞仪检测荷瘤小鼠外周血中CD4+,CD8+T淋巴细胞亚群水平。结果:RT-PCR和ELISA检测结果表明,筛选到的小鼠肝癌细胞株MM45T-HIL-6中存在Hyper-IL-6的表达,而MM45T.Li和MM45T-mock不表达。体内成瘤实验结果显示,与MM45T.Li、MM45T-mock相比,MM45T-HIL-6的成瘤性下降;流式细胞分析结果显示,与接种MM45T.Li和MM45T-mock的小鼠相比,接种MM45T-HIL-6稳定表达HYPER-IL-6基因的小鼠肝癌细胞株的小鼠外周血中CD4+,CD8+T淋巴细胞水平明显升高,差异有统计学意义(P<0.05)。结论:转染Hyper-IL-6基因的小鼠肝癌细胞株能够诱导小鼠抗肝癌主动免疫反应。AIM：To prepare the mouse hepatocellular carcinoma（HCC） cells with the stable expression of Hyper-IL-6 and explore the possibility of inducing active anti-HCC immune response by Hyper-IL-6 gene.METHODS： Hyper-IL-6 gene was transfected into mouse HCC cells MM45T.Li using LipofectamineTM2000.G418-resistant clones named MM45T-HIL-6 were selected and detected for the expression of Hyper-IL-6 gene by RT-PCR and ELISA.Mouse HCC cells transfected with pEGFP-C1,named MM45T-mock,were prepared as controls.Tumor models were established by injecting subcutaneously 5×105 cells of MM45T.Li,MM45T-mock and MM45T-HIL-6 on the right anterior limb of BALB/c mouse,respectively.In vivo experiments were performed to observe the tumorigenicity of MM45T.Li,MM45T-mock and MM45T-HIL-6.The levels of CD4＋ and CD8＋ T cells in mouse peripheral blood were detected by flow cytometry（FCM）.RESULTS： RT-PCR and ELISA showed that Hyper-IL-6 gene was expressed in the MM45T-HIL-6 cells,but not in the control cells.We observed that the tumorigenicity of MM45T-HIL-6 decreased compared with control cells after they were inoculated subcutaneously into mice.FCM results indicated that the levels of CD4＋ and CD8＋ T cells in the peripheral blood significantly increased in the mice inoculated with MM45T-HIL-6 compared with the ones inoculated with MM45T.Li and MM45T-mock cells（P〈0.05）.CONCLUSION： The mouse HCC cells with the stable expression of Hyper-IL-6 can induce active anti-HCC immune response after inoculated subcutaneously into mice.

关 键 词：IL-6 HYPER-IL-6 肝肿瘤 癌症疫苗 抗肿瘤免疫 

分 类 号：R392.11[医药卫生—免疫学] R735.7[医药卫生—基础医学]