PTEN基因重组腺病毒载体对脑胶质瘤U251生长和凋亡的影响

摘要

Objective To investigate the effects of PTEN (phosphatase and tensin homologue deleted on chromosome 10) on the growth and apoptosis of glioma cell line U251. Methods The mRNA and protein expressions of PTEN in glioma cell lines transfected by Ad-PTEN were determined by RT-PCR and Western blotting. Apoptosis of cells were detected by MTT. The alterations of p-Akt and P65 protein levels were examined by Western blotting. Results The success of Ad-PTEN transfection were conifrmed by Fluorescence microscopy Western blot and RT-PCR showed apoptosis of glioma cell lines were significantly higher after transfected by Ad-PTEN. The protein expressions of p-Akt and P65 decreased signiifcantly after transfected by Ad-PTEN. Conclusion Overexpression of PTEN will inhabit the growth through promoting cell apoptosis by inhibiting the expression of p-Akt and NF-кB on glioma cell lines.%目的：探讨PTEN基因对人脑胶质瘤细胞株U251生长及凋亡的影响。方法携带PTEN基因的重组腺病毒载体转染U251细胞株，采用RT-PCR技术和Western blot方法检测Ad-PTEN-GFP组及Ad-GFP及Control组中PTEN mRNA及蛋白水平表达，用MTT及流式细胞仪检测各组细胞生长、周期及凋亡率等变化，Western blot检测细胞蛋白水平表达的变化。结果Ad-PTEN-GFP转染U251细胞后，经荧光显微镜、PCR和Western blot证实转染成功。Ad-PTEN-GFP转染U251细胞后，细胞生长明显受抑制（P<0.05）。Ad-PTEN-GFP转染组中p-Akt及P65蛋白水平下降，与Control组相比差异有统计学意义。结论PTEN基因转染后能抑制人脑胶质瘤U251的生长，调控细胞周期，可能通过PTEN-Akt-NF-кB促进细胞凋亡。