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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: To explore the in vitro and in vivo therapeutic effects of HSV-tk gene / GCV system on glioma in rats. METHODS: STK plasmids were packaged with PA3 1 7 cells to form PA317tk producing virus cells and producing fake retroviral particles. The NIH3T3 cells were used to measure the virus titer. 96 well plates were inoculated with 2 4 well C6 cells and 24 well C6tk + cells. After 48 hours, the medium was changed into culture medium containing GCV. The concentration of GCV was 0,0. 25ug / ml, 0.5ug / ml, 0.75ug / ml, 1 ug / ml, 5 ug / ml, 10 ug / ml, 100 ug / ml increments, each concentration of 3 wells, 5 days after cell viability was measured by MTT method. C6 glioma cells were cultured in vitro and injected stereotaxically into the brain of mice. The size and location of glioma were measured by MRI on days 1, 0, 1, 6 and 25, respectively. In the experimental group, 20 μl of the virus suspension was injected into the tumor and injected every other day. In the control group, an equal volume of normal saline was injected. After the intraperitoneal injection of GCV3 (0 mg · kg-1 · day-1) for 12 days, 1 4 days, MRI measurement of glioma size. Results: The glioma cells transfected with tk gene were highly sensitive to GCV. MRI showed that the introduction of tk gene into intracranial gliomas significantly reduced or disappeared after GCV treatment. Conclusion: The HSV-tk gene / GCV system can effectively kill glioma cells.