双皮质素过表达载体的构建及其对大鼠脑胶质瘤细胞迁移能力的影响  

Constrution of DCX overexpression vector and its effect on migration ability of rat glioma cells

作　　者：高玥 姬广全 高殿帅 Abiola Abdulrahman Ayanlaja 

机构地区：[1]徐州医科大学,江苏省221004

出　　处：《江苏医药》2017年第24期1749-1752,F0002,I0001,共6页Jiangsu Medical Journal

基　　金：国家自然科学基金(81372698)

摘　　要：目的观察过表达双皮质素(DCX)基因对大鼠脑胶质瘤细胞迁移能力的影响。方法采用PCR扩增DCX基因片段,构建DCX慢病毒表达载体,感染293T细胞,荧光显微镜下观察绿色荧光蛋白的表达。筛选稳定表达DCX的大鼠脑胶质瘤C6细胞,将C6细胞分为空载体病毒感染组(GV468组)和过表达DCX病毒感染组(GV468-DCX组),qRT-PCR和Western blot法检测DCX表达。Transwell实验检测过表达DCX对大鼠胶质瘤C6细胞迁移能力的影响。结果成功构建过表达DCX的慢病毒载体,感染C6细胞的效率达到90%。感染C6细胞后,GV468-DCX组DCX mRNA和蛋白表达较GV468组高(P<0.01或P<0.05),GV468-DCX组迁移能力高于GV468组(P<0.05)。结论成功构建DCX慢病毒表达载体,获得稳定表达DCX的大鼠胶质瘤细胞系,过表达DCX基因能促进胶质瘤细胞的迁移能力,为脑胶质瘤的基因治疗提供参考。Objective To observe the influence of doublecortex（DCX）overexpression vector on the migration ability of rat glioma cells.Methods The DCX gene fragment was amplified by PCR and the DCX lentiviral vector was constructed,which then was infected into 293 Tcells.The expression of green fluorescent protein was observed by fluorescence microscope.The rat brain glioma C6 cells were screened for stable expression of DCX,which were divided into GV468 cells（virus-infected empty vectors）and GV468-DCX cells（virus-infected overexpressed-DCX）.The mRNA and protein expressions of DCX were detected by qRT-PCR and Western blot.Transwell assay was used to detect the effect of DCX gene on the migration of C6 cells.Results The lentiviral vector carrying overexpressed GV468-DCX was constructed successfully and the infection efficiency in C6 cells was90%.The mRNA and protein expressions of DCX and the migration ability were higher in GV468-DCX cells than those in GV468 cells（P〈0.01 or P〈0.05）.Conclusion DCX lentiviral expression vector is successfully constructed and the rat glioma cell line stably expressing DCX is obtained.The overexpression of DCX gene enhances the migration ability of glioma cells,which provides a reference for the gene therapy of glioma.

关 键 词：双皮质素 脑胶质瘤 慢病毒 

分 类 号：R739[医药卫生—肿瘤]