白细胞介素12基因与HSV TK基因联合治疗小鼠肝癌的研究  

Combination therapy of murine liver cancer with IL 12 gene and HSV TK gene

作　　者：唐展云[1] 孙文长[1] 陈诗书[1] 

机构地区：[1]上海第二医科大学人类基因治疗研究中心,200025

出　　处：《中华肿瘤杂志》1999年第5期332-335,共4页Chinese Journal of Oncology

基　　金：国家自然科学基金

摘　　要：目的　观察白介素１２（ＩＬ１２） 基因与单纯疱疹病毒胸苷激酶（ＨＳＶＴＫ） 基因联合治疗小鼠肝癌的疗效。方法　将ＩＬ１２ 基因和ＨＳＶＴＫ 基因分别转导入小鼠肝癌ＭＭ４５Ｔ－Ｌｉ 细胞，获稳定表达的ＭＭ４５Ｔ－ＬｉＩＬ１２ 和ＭＭ４５Ｔ－ＬｉＴＫ。于Ｂａｌｂｃ 小鼠皮下接种ＭＭ４５Ｔ－Ｌｉ 细胞２ ×１０５ ，待肿瘤长至０－５～１－０ ｃｍ 时，将ＭＭ４５Ｔ．ＬｉＴＫ细胞与经６０ Ｃｏ 照射的ＭＭ４５Ｔ．ＬｉＩＬ１２ 细胞混合，行瘤内注射治疗，于治疗次日，腹腔注射Ｇａｎｃｉｃｌｏｖｉｒ（ＧＣＶ４０ ｍｇ·ｋｇ－ １·ｄ－ １） ，连续注射１０ 天。按同样方法观察对远侧接种的肿瘤的治疗作用，并通过细胞毒Ｔ淋巴细胞（ＣＴＬ） 活性检测及免疫组化染色，探讨其抗肿瘤机制。结果　ＭＭ４５Ｔ．ＬｉＩＬ１２ 与ＨＳＶＴＫＧＣＶ 联合治疗，肿瘤体积显著缩小，生长受到抑制，疗效显著优于单独治疗组（ Ｐ＜ ０．０１） 。有６０％ 小鼠的肿瘤完全消退，且观察２ 个月无肿瘤生长。两者联合治疗，对远侧肿瘤也具有明显的抗肿瘤作用（ Ｐ＜０．０５） 。ＭＭ４５Ｔ．ＬｉＩＬ１２Objective To investigate the synergistic antitumor effects of murine IL 12 gene and HSV TK gene therapy in mice bearing liver cancer. Methods Mouse liver cancer MM45T. Li (H 2 d) cells were transfected with retroviral vector containing IL 12 gene or HSV/TK gene insert. Gene modified liver cancer cells, MM45T. Li/IL 12 and MM45T. Li/TK, with stable expression of IL 12 and TK were obtained. Balb/c mice were inoculated subcutaneousoly with 2×10 5 MM45T. Li cells. When the tumor reached a size of 0.5～1.0 cm, a mixture of MM45T. Li/TK cells and 60 Co irradiated MM45T. Li/IL 12 cell were injected intratumoraly. Ganciclovir (GCV) was injected ip (40mg·kg -1 ·d -1 ) for 10 days. Intratumoral injection of 60 Co irradiated MM45T. Li/IL 12 cells was repeated twice in one week apart. Mice with distant tumors were treated according to the same protocol. CTL activity of spleen cells was measured by 51 Cr release assay and phenotype of tumor infiltrating lymphocytes by immunohistochemical staining. Results In mice treated with MM45T. Li/IL 12 or MM45T. Li/TK+GCV individiually led to moderate reduction in tumor growth, but neither could eradicate the tumor completely, while in 60% of mice treated with a mixture of MM45T. Li/IL 12 and MM45T. Li/TK cells plus GCV, complete tumor regression was observed, with no tumor recurrence for two months. The growth of distant tumor was also inhibited significanty in mice similarly treated. Most of the mice received combined gene therapy plus GCV had abundant CD4 +, CD8 + T lymphocyte infiltration. Their CTL activity was significantly higher than in mice received single gene therapy. Conclusion Combination therapy with IL 12 gene and HSV TK gene+GCV is effective for mouse liver cancer.

关 键 词：肝肿瘤 治疗 白细胞介素-12 HSVTK基因 基因治疗 

分 类 号：R735.705[医药卫生—肿瘤]