携带白细胞介素12基因逆转录病毒载体包装细胞株体内直接注射治疗肝癌的实验研究

目的研究体内直接注射携带白细胞介素12（mIL-12）逆转录病毒载体包装细胞株对体内肝癌细胞生长抑制作用，探索基因治疗新途径。方法构建携带mIL-12的逆转录病毒载体，将载体转染包装细胞株，用该细胞株分别在大鼠腹腔内与肝癌细胞株混合接种，在肝癌局部及脾内直接注射，观察对肝癌细胞生长抑制作用及其对大鼠毒性反应、免疫功能变化。结果应用该细胞株与肝癌细胞株在腹腔进行混合接种，1：10以下治疗组可使大鼠长生存，而大于1：10治疗组则只能延长生存期。肝癌局部和脾内治疗组中第1天、第3天治疗能使大鼠长期生存，第5天、第7天治疗组则生存期明显延长，二者疗效无明显差别，但脾内治疗组副作用明显减少。免疫学分析与对照组相比，治疗组肝癌组织中浸润的淋巴细胞明显增多，以NK细胞及细胞毒T细胞为主。病理学结果治疗组肿瘤组织血管壁明显增厚，管腔狭窄。结论体内直接注射携带白细胞介素12的逆转录病毒包装细胞株可抑制肝癌细胞生长，早期治疗优于晚期治疗，脾内直接注射治疗是一条安全且有效的肝癌基因治疗的新途径。

Experimental study of the anti-tumor effects of directly administered packaging cell line of retroviral vector carrying IL-12 gene on hepatoma

Objective To study the inhibitory effects of vivo derectly administered pekaging cell line of retroviral vector carrying mouse interleukin-12 (mIL-12 ) on the growth of the vivo established hepatoma and to explore a new way for hepatoma gene therapy. Methods Retroviral vector carrying mIL-12 gene was constructed.The vector was transfected to retroviral peckaging cell line. Then the cell was injected intra-peritoneally and locally into hepatoma tissues or intrasplenally to observe its effects on the growth of hepatoma and the changes of immunological function in rats. its toxic effects were also documented. Results In treatment group, the rats survived permanently with the ration of the packaging cell to hepatoma cell being less than 1: 10, while the survival bine of cd was only prolonged when the ratio was more than 1: 10. In groups treatd locally in the hepatoma tissue or intrasplenally, the rats surived permanantly with the packaging cell line given on the first or the third day, while the rats survived only longer with the packaging cell line given on the fifith or the seventh day. The immunological study showed that the number of hepatoma infiltaing lymphocytes, mainly composed of NK cell and cytotoxic T cell, was increased in the treatment group. The pathological study showal thicking of the vascular wall and stemosis of vascular cavity in hepatoma tissue in the treatmnet group. Conclusion The directly injected packaging cell line in vivo carrying mIl-12 gene via retroviral vector inhibits the growth of hepatoma.The therapeutical effects of early administration are superior to later one Direct to later one. Direct injection via spleen is a new, safe and effective way for hepatoma therapy with.