白细胞介素12及白细胞介素2对大鼠肝癌的联合基因治疗研究

Combined Interleukin 12 and Interleukin 2 Gene Therapy for Hepatocellular Carcinoma in A Rat Model

目的:研究利用小鼠白细胞介素12(mIL-12)和人白细胞介素2(hIL-2)基因对大鼠肝癌进行联合基因治疗的可行性和疗效。方法:构建携带mIL-12和hIL-2基因的逆转录病毒载体,转染包装细胞后对实验性肝癌大鼠进行肝癌局部注射,观察对肝癌细胞的生长抑制作用及其对大鼠的免疫功能变化、毒性反应。结果:携带mIL-12/hIL-2基因的重组逆转录病毒在肿瘤内局部注射明显抑制了肝肿瘤的生长。肝癌接种后第1,3,5,7天治疗组大鼠35d生存率分别为100％,100％,30％,10％。IL-12+IL-2治疗组平均生存时间明显高于生理盐水对照组(P<0.01)、逆转录病毒空载体对照组(P<0.01)、IL-2治疗组(P<0.01)和IL-12治疗组(P<0.05)。治疗后肝癌组织中浸润的淋巴细胞明显增多。结论:肝癌局部注射携带mIL-12和hIL-2基因的逆转录包装细胞株可明显抑制肝癌细胞的生长,早期治疗优于晚期治疗。

Objective:To determine the feasibility and efficacy of combined murine interleukin 12 (mIL-12) and human interleukin 2 ( hIL-2) gene therapy for hepatocellular carcinoma in a rat model. Methods; The retroviral vector encoding mIL-12/hIL-2 gene was constructed and then transfected into packaging cell line. The cells were injected into rats in the established hepatoma at different time points. The therapeutic effect, immune function and toxicological response were evaluated. Results:Intratumoral injection of recombinant retroviral vector encoding mIL-12/hIL-2 gene resulted in marked hepatoma regression. The 35 d survival rates of the subgroups treated on the first,third,fifth and seventh day after tumor implantation were 100% , 100% , 30% and 10% respectively. The average survival time of the IL-12 + IL-2 treatment group was superior to those of the physiological saline group (P <0. 01) , retroviral empty vector group(P<0.01 ) , IL-2 treatment group(P <0. 01) and IL-12 treatment group( P <0. 01). The immunological study showed that the number of hepatoma infiltrating lymphocytes was increased in the IL-12 + IL-2 treatment group. Conclusion; The retroviral packaging cell line encoding mIL-12 and hIL-2 gene via intratumoral injection inhibits the growth of hepatocellular carcinoma significantly. The therapeutical effects of early administration is superior to that of later one.