摘要
目的探讨瘤内注射表达hIL-2质粒DNA抗小鼠肝癌皮下移植瘤的作用。方法构建真核表达质粒载体pDC511hIL-2;ELISA方法检测质粒载体在真核细胞中的表达;胸腺细胞增殖法(MTT法)检测hIL-2生物学活性。小鼠肝癌H22皮下移植瘤瘤内直接注射质粒DNA后,观察各组小鼠存活时间,肿瘤体积变化;检测小鼠脾脏细胞毒T淋巴细胞(CTL)杀伤活性。在质粒DNA注射后一月进行瘤体组织病理学观察。结果构建的质粒载体能在真核细胞内高效表达hIL-2(3087.18pg/ml.24h.106)。质粒载体表达的hIL-2体外刺激胸腺细胞增殖作用非常明显。hIL-2基因治疗组与空载体组相比,肿瘤生长明显受抑制(F=3.85,P=0.03),小鼠存活期显著延长(x2=4.10,P=0.03),并且小鼠脾细胞CTL杀伤活性增强。pDC511hIL-2DNA瘤内注射一月后,肿瘤病灶炎性细胞浸润明显,病灶内肿瘤细胞广泛坏死。结论瘤内注射hIL-2表达质粒DNA可抑制小鼠肝癌皮下移植瘤的生长,提高荷瘤生存期。
Objective To explore the antitumor efficacy of intra-tumoral administration of plasmid DNA expressing hIL-2 in murine tumor models.Methods Plasmid encoding hII2 was constructed and cloned. The expression of the cytokine in the eukaryotic cells was assessed by enzyme-linked immunosorbet assay. The proliferation assay of thymocytes was performed for measuring the biological activity of expressed hIL2. After intra-tumoral administration of plasmid DNA, mean diameters of the tumor mass and survival time were measured in each murine models. CTL cytolytic activity was performed with different plasmid DNA for its action against H22 cells. Histopathological analysis was operated after administration of plasmid DNA vectors in murine model group. Results Plasmid vector could efficiently express hIL-2 （3087.18pg/ml·24h·10^6 cell ）, which could significandy stimulate the proliferation of thymocytes in vitro. Growth of tumor was significantly inhibited（F= 3.85,P = 0.03）, and activity of CTL against H22 cell was enhanced in hIL2 gene therapy group as compared with the control group. In the foci treated with pDCSllhIL2 plasmid DNA, inflammatory cell infiltration was more extensive and necrosis was more definite than control group at 1 month after DNA injection. Conclusion Intra-tumoral administration of plasmid DNA encoding interleukin-2 can inhibit the growth of transplanted tumor and induce the host antitumor immune response efficiently in the murine model.
出处
《实用肝脏病杂志》
CAS
2005年第5期257-260,共4页
Journal of Practical Hepatology
