IL-2和IL-12联合基因治疗小鼠肝癌的实验研究  

Gene therapy with IL-2 and IL-12 for murine hepatocellular carcinoma

作　　者：刘勇[1] 胡大荣[1] 谭晓华[1] 胡学玲[1] 范公忍[1] 韩聚强[1] 吴忆贫[1] 

机构地区：[1]北京军区总医院全军肝病治疗中心,北京100700

出　　处：《第三军医大学学报》2004年第12期1071-1074,共4页Journal of Third Military Medical University

摘　　要：目的　探讨瘤内注射共表达mIL 12和hIL 2质粒DNA抗小鼠肝癌皮下移植瘤的作用。方法　构建pDC5 11hIL2 mIL12、pDC5 11mIL 12、pDC5 11hIL 2真核表达质粒载体 ;ELISA方法检测各组质粒在真核细胞的表达 ;小鼠肝癌H2 2皮下移植瘤瘤内注射各组质粒DNA后 ,检测不同时间血清中细胞因子浓度 ;观察各组小鼠存活时间 ,肿瘤大小变化 ;并检测各组小鼠脾脏细胞毒T淋巴细胞 (cytotoxicTlymphocyte ,CTL)活性。对各治疗组在质粒DNA注射后一月进行瘤体组织学观察。结果　酶切鉴定各组质粒载体 ,均示构建成功 ,并能在真核细胞内高效表达相应细胞因子。瘤内注射各组质粒载体后 ,pDChIL2 mIL12组在不同时间表达的hIL2和mIL12分别与pDC5 11hIL2组 (F =71 11,P <0 0 1)、pDC5 11mIL12组 (F =3 0 70 ,P <0 0 5 )比较有显著性差异。mIL 12基因和hIL 2基因联合治疗组 ,肿瘤生长明显受抑制 ,疗效显著优于各单独治疗组和对照组 (P <0 0 5 ) ,并且小鼠脾细胞CTL杀伤活性增强。双基因联合治疗组 ,病灶内肿瘤细胞坏死明显 ,炎性细胞广泛浸润。结论　IL 12、IL 2基因治疗可抑制小鼠肝癌H2 2皮下移植瘤的生长 ,提高机体的抗肿瘤免疫应答 ,两者联合运用可产生协同效应。Objective To explore the anti tumor efficacy of intratumor administration of plasmid DNA expres sing both mIL12 and hIL2 in mice H22 hepatocellular carcinoma grafted subcutaneously. Methods Eukaryotic expression plasmid vectors of hIL2 mIL12, mIL12, and hIL2 were constructed. The expressions of the plasmid in the the eukaryotic cells in different groups were examined by enzyme linked immunosorbent assay (ELISA). Cytokine expression in serum was detected at different time after intratumor administration of plasmid DNA. The mean diameter of the tumor mass and the livability of mice were measured in each mouse model group. Lactic dehydrogenase (LDH) assay was used to examine whether or not the treatment with different plasmid DNA could induce systemic cytolytic activity of lymphocytes against parental H22 cells. Histopathological changes were observed after administration of plasmid DNA vectors in each mouse model group. Results Plasmid vectors were constructed successfully and could efficiently express cytokines. After the intratumor administration, the expression levels of hIL2 and mIL12 in the pDC511hIL2 mIL12 group were statistically different as compared with that in group pDC511hIL2 ( F =71.11, P <0.01) or group pDC511mIL12 ( F =30.70, P <0.05) at different time, respectively. Growth of tumor in combined gene therapy group was significantly inhibited as compared with that in other groups ( P <0.05). Enhanced activity of CTL was observed in the pDC511hIL2 mIL12 group. In the group treated with pDC511hIL2 mIL12 plasmid DNA, inflammatory cell infiltration was more extensive and necrosis was more definite than those in other groups. Conclusion Both IL 2 and IL 12 are able to inhibit the growth of mice H22 hepatocellular carcinoma grafted subcutaneously and induce the host anti tumor immune response efficiently. Combination of IL 2 and IL 12 may have synergistic effect in anti tumor.

关 键 词：肝细胞癌 MIL-12 HIL-2 免疫基因治疗 动物模型 

分 类 号：R392.11[医药卫生—免疫学] R730.59[医药卫生—基础医学]