摘要
目的将携带有PTEN基因的重组腺病毒转染大鼠肾脏成纤维细胞,观察其过度表达PTEN对细胞周期的影响。方法以免疫组化的方法确定PTEN在成纤维细胞中的亚细胞定位,以Ad-PTEN转染成纤维细胞,荧光倒置显微镜下观察绿色荧光蛋白表达情况及亚细胞定位,将转染的成纤维细胞及正常对数生长期成纤维细胞消化,流式细胞仪分别检测细胞周期。结果免疫组化结果显示成纤维细胞的胞浆和胞核中均有PTEN的表达,在胞核中表达水平更高,荧光倒置显微镜下观察Ad-PTEN转染成纤维细胞后外源性PTEN其标志物绿色荧光蛋白(GFP)在胞浆和胞核中均表达,其中胞核表达水平更高,流式细胞仪检测在成纤维细胞中病毒转染组G1/S均明显高于对照组,提示Ad-PTEN转染组细胞存在明显的G1期细胞阻滞。结论PTEN过度表达将引起肾脏成纤维细胞周期G1期阻滞,这一过程与PTEN在细胞核定位有关。
Objective To observe the influence of overexpression of PTEN by transfection of recombinant adnovirus containing wild-type PTEN on cell cycle of rat renal fibroblasts. Methods Sub-cellular location of PTEN was identified by immunochemistry; Sub-cellular location of GFP, Mark protein of PTEN was observed through fluorinverted microscope; cell cycle was idendifed by flow cytometry between transfected fibroblast cells and control cells. Results There were PTEN expresses both in cytoplasm and nuclei in fibroblast cells, with a preferential nuclear localization . So as the exogenous PTEN represented by GFP. G1 arrest was observed in the transfected cells by flow cytometry. Conclusion Overexpression of PTEN will induce G1 arrest in rar renal fibroblast cells,which associated with the nuclear location of PTEN.
出处
《重庆医学》
CAS
CSCD
2007年第5期409-410,共2页
Chongqing medicine
