Screening glioma stem cells in U251 cells based on the P1 promoter of the CD133 gene

摘要: Cluster of differentiation (CD)133 is an important cell surface marker glioma stem cells (GSCs). The transcription the CD133 gene controlled by five alternative promoters (P1, P2, P3, P4 and P5), which are expressed in a tissue-specific manner. In present study, recombination technology was used to construct two types expression vectors that contained P1 promoter gene, regulated either neomycin-resistance or herpes simplex virus thymidine kinase (HSV-TK) gene. Following stable transfection U251 glioblastoma with these vectors, expressing were named (+) cells, while regulating HSV-TK called (-) cells. detected flow cytometry reverse transcription-quantitative polymerase chain reaction. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay assess proliferation ability, cycle analyzed cytometry, clone formation test performed evaluate invasive capability results demonstrated that, due expression, ability significantly higher than those conclusion, study successfully established novel method screening GSCs based on