Transcription profiling by array of human PTEN null U87MG glioblastoma cells transfected with wild-type PTEN, mutant PTEN or GFP controls to investigate the effects of PTEN activity on gene expression

To investigate the effects of PTEN activity on gene expression in 3D cultured glioblastoma cells and the contributions of PTEN's lipid and protein phosphatase activities to these effects. We expressed wild type PTEN, a catalytically dead mutant (C124S) or mutants selectively lacking protein or lipid phosphatase activity (Y138L and G129E respectively) in U87MG cells at physiological levels by transient unselected polyclonal lentiviral transduction. Cells were then seeded into 3D matrigel and RNA prepared after 15 hours.

本研究旨在探讨磷酸酶与张力蛋白同源物（PTEN）活性对三维培养胶质母细胞瘤细胞基因表达的影响，以及PTEN的脂质磷酸酶和蛋白磷酸酶活性对上述效应的具体贡献。我们通过未经筛选的瞬时多克隆慢病毒转导，在U87MG细胞中以生理水平表达野生型PTEN、催化失活突变体（C124S），以及分别选择性缺失蛋白磷酸酶活性（Y138L）和脂质磷酸酶活性（G129E）的突变体。随后将细胞接种至三维基质胶（Matrigel）中，15小时后提取RNA。