3种不同类型基因导入系统转染类神经元细胞的效率比较  

The comparison of the transfection efficiency of three different types of gene expression system in the neurotypic cell lines

作　　者：梁兵[1] 袁芳[1] 杨宁[2] 殷建瑞[1] 蒲蜀湘[1] 解龙昌[1] 高庆春[1] 高聪[1] 

机构地区：[1]广州医学院第二附属医院神经科学研究所/神经内科,广州510260 [2]广州医学院第五附属医院神经内科,广州510700

出　　处：《重庆医学》2012年第18期1792-1794,1798,共4页Chongqing medicine

基　　金：国家自然科学基金青年项目(81100890);广东省自然科学基金资助项目(10151018201000022);广东省博士启动项目(10451018201004383);广州市属高校羊城学者科研项目(10A010G);广州医学院博士启动基金(2008c46)

摘　　要：目的应用3种不同类型基因导入系统携带能表达绿色荧光蛋白的pAAV-EGFP质粒转染两类神经元细胞株,并对细胞毒性和转染效果进行比较。方法 LipofectAMINE 2000、聚乙烯亚胺(polyethylenimine,PEI,相对分子质量为25×103)及该课题组合成的PEI-聚乙二醇(polyethyleneglycol,PEG)4.6在SH-SY5Y神经母细胞瘤细胞和C6胶质瘤细胞中的转染效果,通过MTT法检测细胞存活率,倒置显微镜荧光观察法和流式细胞仪法检测转染效果。结果 MTT检测发现,在C6胶质瘤细胞中,PEG-PEI 4.6的细胞存活率为83%,比PEI(70%)和LipofectAMINE 2000(73%)的细胞存活率要高(P<0.05);但Lipo-fectAMINE 2000和PEI的细胞存活率比较,差异无统计学意义(P>0.05);在SH-SY5Y神经母细胞瘤细胞中的检测结果与C6胶质瘤细胞类似。倒置显微镜观察发现,在C6胶质瘤细胞中,PEG-PEI 4.6的细胞荧光表达效果比LipofectAMINE2000和PEI要好;而在SH-SY5Y神经母细胞瘤细胞中,LipofectAMINE 2000的荧光表达效果比另外两种复合物的效果好,同时PEG-PEI 4.6的荧光表达效果比PEI要好。流式细胞仪检测发现在C6胶质瘤细胞中,PEG-PEI 4.6的转染效率最好,为23.2%;Lipo-fectAMINE2000其次,为16.9%;PEI最差,为12.6%,三者两两比较差异有统计学意义(P<0.05)。在SH-SY5Y神经母细胞瘤细胞中则是LipofectAMINE 2000的转染效率最好,PEG-PEI 4.6其次,PEI最差,转染效率分别为22.3%、17.2%、10.6%,三者两两比较差异有统计学意义(P<0.05)。结论 LipofectAMINE 2000和PEG-PEI 4.6都可作为目前神经系统基因转染的传输载体。Objective Use three different types of gene expression system carry the pAAV-EGFP plasmid can express green flu- orescent protein to transfect the neurotypic cell lines and to compare their cell toxicity and their transfection effects. Methods We used the commercial liposomes and nanoparticles such as the LipofectAMINE 2000,the PEI（25×10^3） and PEG-PEI 4. 6 as the gene expression system,carry the pAAV-EGFP plasmid to transfect the commonly used neurotypic cells in nervous system cells such as the SH-SY5Y neuroblastoma cells and C6 glioma cells. Then we detected the cell toxicity through the MTT method, and de- tected the transfection effects by the inverted microscope fluorescent observation and the flow cytometric method. Results MTT detection results showed the cell survival rate of PEG-PEI 4.6 （83%）was higher than PEI（70 % ）and LipofectAMINE 2000（73%） in the C6 glioma cells （P〈0.05） ,but the compare of the cell survival rate of PEI and LipofectAMINE 2000 had no statistical sig- nificance（P^0.05） ;The similar results were in the SH-SY5Y neuroblastoma cells. The inverted microscope fluorescent observed in C6 glioma cells the PEG-PEI 4.6 had higher fluorescence expression than LipofectAMINE2000 and PEI. While in SH-SY5Y neuro- blastoma cells,the LipofectAMINE2000 had higher fluorescence expression than other two materials. At the same time, the PEG- PEI 4.6 had higher fluorescence expression than PEI. The flow cytometric results showed in （;6 glioma cells the PEG-PEI 4.6 had the best efficiency to 23.2%, LipofectAMINE2000 was next of 16.9%, the worst PEI was 12.6 %（P〈0.05） In SH-SY5Y neuro- blastoma cells LipofectAMINE2000 had the best efficiency,PEG-PEI 4.6 second, and PEI was the worst （P〈0.05）, the efficiency of them was 22.3 %, 17.2 % and 10.6% respectively. Conclusion At present LipofectAMINE 2000 and PEG-PEI 4.6 can serve as the gene transfection carrier of the nervous system.

关 键 词：神经胶质瘤 神经母细胞瘤 脂质体 转染 

分 类 号：R440[医药卫生—诊断学]