逆转录病毒介导Fcy∷Fur/5-FC基因治疗对脑胶质瘤杀伤作用的研究  

Effects of retrovirus-mediated Fcy∷Fur/5-FC gene therapy for glioma

作　　者：封林森[1] 胡卫星[1] 卢春[2] 唐桂霞[2] 史德志[1] 贾雪梅[2] 

机构地区：[1]南京医科大学第一附属医院神经外科,江苏南京210029 [2]南京医科大学微生物免疫教研室,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2006年第1期47-50,共4页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省医学重点人才"135"工程资助项目(苏卫科教2001-31)

摘　　要：目的:研究融合型自杀基因Fcy∷Fur联合5-FC对胶质瘤细胞株C6的杀伤效果。方法:扩增Fcy∷Fur基因并构建Fcy∷Fur基因重组逆转录病毒载体PLXSN-Fcy∷Fur;载体转染包装细胞PT67获得高滴度病毒再转染鼠胶质瘤细胞C6,筛选并鉴定阳性转基因克隆;以MTT法和FCM法检测5-FC对Fcy∷Fur转基因细胞的杀伤效应。结果:PCR法扩增出全长Fcy∷Fur基因,经测序证实序列正确;PLXSN-Fcy∷Fur滴度为3×106CFU/ml;RT-PCR检测显示Fcy∷Fur转基因阳性克隆的C6细胞有效表达目的基因的mRNA;应用5-FC后FCM法检测到显著的凋亡峰,MTT法检测细胞有明显的杀伤效应。结论:融合型自杀基因Fcy∷Fur联合5-FC可对胶质瘤细胞C6产生明显的杀伤作用。Objective： To investigate the killing effects of Fcy ：： Fur suicide gene therapy system on rat malignant glioma cells C6 in vitro. Methods： Fcy ：： Fur gene was cloned and the retroviral vector Fcy ：： Fur was constructed. The vector was transfered into packaging cell line PT67 and high-titer virus was abtained to infect the tumorigenic glioma cell line C6. Finally, the killing efficacy of 5-FC on the gene-transfered cells was determined by FCM and MTT assays. Results： Fcy ：： Fur gene was cloned and its sequence was confirmed. A retroviral vector of Fcy ：： Fur, PLXSN- Fcy ：： Fur, was constructed and transfered into packaging cell line PT67. Hightiter（3×10^6 CFU/ml）virus was obtained. The FCM and MTT assays showed 5-FC killed C6-Fcy ：： Fur cell at the concentration of 50μg/ml. Conclusion： Fusion suicide gene therapy system Fcy ： ： Fur/5-FC has a significant killing efficacy on gene-transfered cell C6.

关 键 词：融合型自杀基因 Fcy::Fur 逆转录病毒 胶质瘤细胞 C6 

分 类 号：R739.41[医药卫生—肿瘤]