纳米材料PEG-PEI介导双基因促体外拟神经细胞凋亡的作用

Nano-material mediated combined genes to promote neural-type cells apoptosis in vitro

目的探讨新型纳米材料PEG-PEI介导的双基因pIRES2-EGFP/CD-5-FC和pIRES2-EGFP/TRAIL在体外对拟神经细胞模型(神经母细胞瘤株SH-SY5Y细胞)的联合杀伤作用。方法将PEG-PEI介导的联合基因pIRES2-EGFP/CD-5-FC和pIRES2-EGFP/TRAIL转染SH-SY5Y细胞后,采用MTT法观察其对SH-SY5Y细胞的杀伤作用,荧光显微镜下观测以及流式细胞仪检测SH-SY5Y细胞凋亡率及FPEG-PEI的转染效率。结果 N/P=15时PEG-PEI转染两种目的基因中单一基因的细胞凋亡作用最强(P<0.01);两种基因联合转染SH-SY5Y细胞时的细胞凋亡率为77%,较单一基因转染的细胞凋亡率提高了27%(P<0.01)。结论 CD-5-FC和TRAIL基因在体外联合转染对SH-SY5Y细胞的杀伤作用较单一基因更强;PEG-PEI可以作为神经细胞基因靶向传输的载体行神经系统疾病基因治疗的体内实验研究。

Objective To evaluate transfection efficiency of PEG-PEI and the antitumor effect when the CD and TRAIL genes were jointly used against human neuroblastoma cells （SH-SYSY） in vitro. Methods The SH-SY5Y ceils were treated by the combined application of PEG-PEI/pIRES2-EGFP/CD and PEG-PEI/pIRES2-EGFP/TRAIL. The trans- fection efficiency of PEG-PEI and the antitumor effect were evaluated by MTY assay, fluorescence microscope and flow cy- tometry assay. Results This study revealed a significant increase of antitumor effect in vitro following the combined appli- cation of PEG-PEI/pIRES2-EGFP/CD and PEG-PEI/pIRES2-EGFP/TRAIL treatments in SH-SYSY cells, and the lethal effect by united transfection was stronger than that by single transfection. Conclusions United gene transferation has more stronger lethal effect than single gene. PEG-PEI acts as targered gene transmission carrier of nerve cell for empirical study of gene therapy in nervous system disease in vivo.