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为了研究自杀基因——胞嘧啶脱氨酶基因(cytosine deaminase gene,CD基因)对大鼠脑胶质瘤的体内治疗效果,采用基因转导系统将慢病毒包装的CD基因转染骨髓间充质干细胞(MSC)并使其长时间、高效表达,然后移植到使用颅内立体定向接种法制作的40只SD大鼠胶质瘤模型中.按接种细胞类型将实验SD大鼠分为5组,每组8只:①C6胶质瘤;②C6+MSCs细胞(1∶1);③C6+MSCs细胞(1∶2);④C6+MSC-codA/eGFP细胞(1∶1);⑤C6+MSC-codA/eGFP细胞(1∶2).瘤龄7天后腹腔注射500 mg/(kg.d)5-氟胞嘧啶(5-flucytosine,5-FC),共14天.用磁共振成像(MRI)动态监测肿瘤的体积并进行了大鼠存活期观察、常规病理分析、RT-PCR检测、HE染色.结果显示,第①组瘤龄14天时病灶呈圆形,中心见坏死区,肿瘤平均246 mm3,均存活期15.3天,第②组平均生存期16.0天,第③组平均生存期16.6天,第④⑤组自然存活期均大于30天,14天时病灶平均体积分别为55 mm3、40 mm3,28天时肿瘤抑制率分别为77.24%、83.28%.MRI扫描可清楚显示肿瘤的大小、形态和内部结构,与病理结果高度相关;MRI动态观察可证实携带CD基因的骨髓间充质干细胞及5-FC治疗系统治疗C6颅内胶质瘤有效;RT-PCR检测结果证实肿瘤组织内有胞嘧啶脱氨酶表达.
In order to study the in vivo therapeutic effect of suicide gene cytosine deaminase gene (CD gene) on glioma in rats, gene delivery system was used to transfect lentivirus-packaged CD gene into bone marrow mesenchymal The stem cells (MSCs) were cultured in vitro for 40 days and then were transplanted into 40 SD rat glioma models by intracranial stereotactic inoculation.The experimental SD rats were divided into 5 groups according to the cell type of inoculation, Each group of 8: ① C6 glioma; ② C6 + MSCs cells (1: 1); ③ C6 + MSCs cells (1: 2); ④ C6 + MSC-codA / eGFP cells eGFP cells (1: 2) were injected intraperitoneally with 500 mg / (kg.d) of 5-flucytosine (5-FC) for 7 days.A total of 14 days were observed with magnetic resonance imaging The tumor size and the survival of rats were observed, routine pathological analysis, RT-PCR detection, HE staining.The results showed that the first group of tumor growth 14 days tumor was round, the center of the necrotic area, the average tumor was 246 mm3, were The survival time was 15.3 days. The average survival time in group ② was 16.0 days. The average survival time in group 3 was 16.6 days. The natural survival time in group ④⑤ was more than 30 days. The average volume of lesions was 55 m m3, 40 mm3, the tumor inhibition rates at 28 days were 77.24% and 83.28% respectively.MRI scan could clearly show the size, shape and internal structure of the tumor, which was highly correlated with the pathological results. MRI dynamic observation confirmed that bone marrow intercellular charge C6 stem cells and 5-FC treatment system for the treatment of intracranial glioma effectively; RT-PCR test results confirm the expression of cytosine deaminase in the tumor tissue.