论文部分内容阅读
目的研究不同拷贝数CArG/5HRE+HSV-TK系统辅助伽玛刀杀伤C6胶质瘤效果,寻求伽玛刀条件下对胶质瘤杀伤更有效的适宜拷贝数CArG/5HRE+HSV-TK系统。方法在Wistar大鼠颅内C6胶质瘤模型的基础上,构建不同拷贝数CArG/5HRE+HSV-TK质粒(空质粒转染+伽玛刀照射组、HSV-TK+伽玛刀照射组、4CArG/5HRE启动子+伽玛刀照射组、9CArG/5HRE启动子+伽玛刀照射组、12CArG/5HRE启动子+伽玛刀照射组),通过脂质体转染到大鼠颅内胶质瘤组织中,对瘤组织伽玛刀放射外科治疗后取大鼠肿瘤组织。通过对瘤组织常规病理、瘤体细胞凋亡、HSV-TK mRNA及蛋白表达的观察,确定不同拷贝数CArG/5HRE+HSV-TK系统辅助伽玛刀诱导HSV-TK表达及对C6胶质瘤杀伤效果。结果各组肿瘤细胞均可见凋亡、破坏,CArG/HRE双启动子+伽马刀照射组更为明显(P<0.05),尤以9CArG/5HRE启动子+伽马刀照射组最为突出,凋亡率为(0.71±0.12)%(P<0.05);CArG/HRE双启动子+伽马刀照射组中HSV-TK mRNA及蛋白表达较无启动子组明显增加(P<0.05),9CArG/5HRE启动子+伽马刀照射组最为明显,HSV-TK mRNA的相对表达量为(0.84±0.09,P<0.05)。结论在SRS治疗条件下,CArG/HRE双启动子能促进目的基因HSV-TK的表达,增加对C6胶质瘤细胞的杀伤,其中,9CArG/5HRE双启动子的作用最明显。
Objective To study the effect of Gamma Knife-killing C6 glioma with different copies of CArG / 5HRE + HSV-TK system and seek for a more effective copy number CArG / 5HRE + HSV-TK system for glioma killing under gamma knife. Methods Based on the intracranial C6 glioma model in Wistar rats, different copy number CArG / 5HRE + HSV-TK plasmids were constructed (empty plasmid transfected + gamma knife irradiation group, HSV-TK + gamma knife irradiation group, 4CArG / 5HRE promoter + gamma knife irradiation group, 9CArG / 5HRE promoter + gamma knife irradiation group, 12CArG / 5HRE promoter + gamma knife irradiation group) were transfected into rat intracranial glioma Tissue, the tumor tissue gamma knife after radiosurgery treatment of rat tumor tissue. The expression of HSV-TK and the expression of HSV-TK mRNA and protein were observed by routine pathology, tumor cell apoptosis and expression of HSV-TK in CArG / 5HRE + HSV-TK system. Killing effect. Results The apoptosis and destruction of tumor cells were observed in all groups, especially in the CArG / HRE dual promoter plus gamma knife irradiation group (P <0.05), especially in the 9CArG / 5HRE promoter + gamma knife irradiation group (0.71 ± 0.12)% (P <0.05). The expression of HSV-TK mRNA and protein in CArG / HRE dual-promoter plus gamma knife group was significantly higher than that in the non-promoter group (P <0.05) The expression of HSV-TK mRNA was the most obvious in 5HRE promoter + gamma knife irradiation group (0.84 ± 0.09, P <0.05). Conclusions The dual promoter of CArG / HRE can promote the expression of HSV-TK gene and increase the killing rate of C6 glioma cells under the treatment of SRS. Among them, the 9CArG / 5HRE double promoter has the most obvious effect.