HSV-tk基因/GCV系统治疗鼠脑胶质瘤实验研究  

Study of retrovirus mediated HSV-tk gene/GCV system therapy for glioma

作　　者：李承晏[1] 李涛[1] 余绍祖[1] 曾庆杏[1] 王镇涛[1] 夏明珠[2] 李茂进[3] 

机构地区：[1]湖北医科大学附一院神经科,430060 [2]湖北医科大学附一院中医科,430060 [3]湖北医科大学附一院放射科,430060

出　　处：《卒中与神经疾病》2000年第1期10-13,共4页Stroke and Nervous Diseases

基　　金：湖北省科委自然科学基金!SJ-97J0 69

摘　　要：目的 :探索 HSV-tk基因 /GCV系统对大鼠脑胶质瘤的体内外治疗效果。方法 :用 PA3 1 7细胞包装STK质粒 ,形成产病毒细胞 PA3 1 7tk,产生假逆转录病毒颗粒 ,NIH3 T3细胞测定病毒滴度。96孔培养板接种 2 4孔C6细胞 ,2 4孔 C6tk+细胞 ,4 8小时后换入含 GCV的培养液 ,GCV浓度按 0、0 .2 5ug/ml、0 .5ug/ml、0 .75ug/ml、1 ug/ml、5ug/ml、1 0 ug/ml、1 0 0 ug/ml递增 ,每种浓度 3孔 ,5天后 MTT法测量细胞存活率。体外培养鼠胶质瘤细胞C6,立体定向注入鼠颅内 ,分别在第 1 0、1 6、2 5天 ,MRI测量胶质瘤大小、位置。实验组瘤内注入 2 0 ul病毒悬液 ,隔日再注入一次 ,对照组注射等体积生理盐水 ,次日起腹腔注射 GCV3 0 mg· kg- 1·day- 1 ,连续 1 2天 ,治疗后第 1 4天 ,MRI测量胶质瘤大小。结果 :转 tk基因后的胶质瘤细胞对 GCV高度敏感。 MRI显示将 tk基因导入颅内胶质瘤 ,GCV治疗后肿瘤明显减小或消失。结论 :HSV-tk基因 /GCV系统能有效杀伤脑胶质瘤细胞。Objective: The study is to evaluate effectiveness of HSV tk gene/GCV system therapy for glioma. Methods: The retroviral vector carrying HSV tk gene was transferred into packaging cell line PA317 to obtain replication defective vector virus. NIH3T3 cells measured virus titers. C6 and C6 tk + cells were cultured in culture medium conclude GCV in different concentration: 0、0.25ug/ml、0.5ug/ml、0.75ug/ml、1ug/ml、5ug/ml、10ug/ml、100ug/ml. The livability of cells was determined by MTT technique five days later. Implanted C6 cells in rats to establish glioma model. MRI measured the size and the site of the glioma. Injected 20ul virus in glioma of the experiment group. The controls were injected by saline instead. Repeated it two days later. 30mg·kg -1 ·day -1 of GCV were intraperitoneally injected for 12 days. MRI measured the size two days later after therapy. Results: In vitro, glioma cells which were transfect by tk gene were very sensitive to GCV. In vivo, MRI observed that after therapy, glioma diminished a lot or disappeared. Conclusion: In vitro and in vivo, HSV tk gene/GCV system can therapy glioma effectively.

关 键 词：HSV-TK基因 基因治疗 脑胶质瘤 

分 类 号：R739.410.5[医药卫生—肿瘤]