中国实用外科杂志

中国实用外科杂志 ›› 2006, Vol. 26 ›› Issue (09): 1-690.

• 论著 •    

IL-2基因转染联合CTL杀伤肝癌细胞的体外试验研究

叶小鸣，左继东，朱易凡   

  1. 中山大学附属第一医院特种医疗中心外科，广东广州510080
  • 收稿日期:2006-04-15 修回日期:2006-06-28 出版日期:2006-08-21 发布日期:2006-08-21

In vitro Studies of cytotoxicity of interleukin2 gene transfection conjugating specific cytotoxic T lymphocyte to hepatocarcinoma cell.

Ye Xiaoming,Zuo Jidon,Zu Yifan.   

  1. Department of Surgery of the Private Medical Centre,the First Affiliated Hospital of SUMS,Guangzhou 510080,China
  • Received:2006-04-15 Revised:2006-06-28 Online:2006-08-21 Published:2006-08-21

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摘要：

目的:通过研究白细胞介素-2（IL-2）基因转染联合特异性细胞毒T淋巴细胞（CTL）对人肝癌细胞（HepG2）的体外杀伤试验，探讨一种肝癌的免疫基因治疗方法。方法:采用多聚阳离子脂质体作载体，将IL-2基因转染至HepG2细胞，通过检测IL-2浓度变化，观察转染效率。用MTT法检测其联合特异性CTL细胞对HepG2细胞的杀伤作用。结果:转染IL-2基因至HepG2细胞的体外最佳转染比例为5∶1，其可持续分泌IL-2 30d，分泌高峰在转染后第2～3天。联合特异性CTL细胞，对HepG2细胞显示明显的协同杀伤效应。结论:经转染IL-2基因后，HepG2细胞分泌IL2可达30d，可以有效地维持特异性CTL细胞的活力，两者联合，对杀伤肝癌细胞有明显的协同效应。

关键词: 白细胞介素2, 细胞毒T淋巴细胞, 免疫基因治疗, 肝癌细胞

Abstract:

Objective:To investigate cytotoxicity of conjugation of interleukin-2 (IL-2) gene transfection and specific cytotoxic T lymphocyte to human hepatacarcinoma cell (HepG2),to seek a new way of immunegene therapy for hepatocarinama. Method:IL-2 gene were transfected to HepG2 cell mediated by polycationic lipid.Efficiency of transfection was observed by detecting changes of culture medium’s IL-2 concentration.Cytotoxicity of conjugation of IL-2 transfection and specific CTL was determined by MTT assay. Result:After transfected IL-2 mediated by polycationic lipid, HepG2 can continuously secrete IL-2 for 30 days;and at 2nd~3rd day,its peak of secretion presence;and at 7th day, its secretion descent obviously.The best transfection ratio is 5∶1 in vitro.IL-2 transfection conjugating specific cytotoxic T lymphocyte can occurrence obvious synergistic effect in cytotoxictiy to HepG2. Conclusion:After transfected IL-2 gene mediated by lipid,HepG2 can continuously secrete IL-2 for 30 days,and this can maintenance activity of specific CTL effectually.The conjugation of IL-2 transfection and specific CTL can show the obvious synergistic killing effect to hepatocarcinoma cell.

Key words: Interleukin2, Cytotoxic T cell, Immunegene therapy, Hepatocarcinoma cell