TDAG51基因在PC12细胞中的过表达及其致凋亡作用  

Overexpression of TDAG51 gene in PC12 cells and its function of inducing apoptosis

作　　者：李宪奎[1] 张百芳[1] 彭芳芳[1] 杞少华[1] 沈晗[1] 吴少波[1] 武栋成[1] 

机构地区：[1]武汉大学医学部生物化学与分子生物学系,湖北武汉430071

出　　处：《新乡医学院学报》2005年第2期102-103,106,F002,共4页Journal of Xinxiang Medical University

摘　　要：目的　探讨TDAG5 1基因在PC12细胞中的表达及其作用机制。方法　实验分为3组,实验组用绿色荧光蛋白质粒(p EGFP - C1 )载体把外源性TDAG5 1基因转染的PC12细胞;空载体对照组用p EGFP- C1 转染的PC12细胞;对照组未转染任何试剂的PC12细胞。在转染后2 4 h、4 8h、72 h收集细胞,用Western blotting检测过表达的TDAG5 1蛋白;转染后84 h用相差显微镜观察PC12细胞形态的变化;Hoechst332 5 8染色观察细胞核形态变化;检测凋亡执行蛋白Caspase- 3的变化。结果　TDAG5 1基因在PC12细胞中高表达;TDAG5 1基因过表达的细胞与两对照组细胞相比较,胞体变圆,轴突缺失;荧光显微镜下观察PC12细胞可见胞核固缩、染色质凝集,并可见核碎片;前Caspase- 3蛋白活化被切割;活化的17k Da Caspase- 3随TDAG5 1蛋白的作用时间延长而增多。结论　过表达TDAG5 1基因可诱导PC12细胞凋亡。Objective To investigate the expression of TDAG51 gene in PC12 cells and its function of inducing apoptosis.Methods The experiment was carried out in three groups.In the experimental group,PC12 cells transfected from TDAG51 gene by pEGFP-C 1 carriers were used;in the non-carrier comparison group,PC12 cells transfected from pEGFP-C 1 were used; in the control group,PC12 cells which were not transfected were used.The above celles were collected at 24 h,48 h and 72 h respectively after transfection.Western blotting was employed to detect the expression of TDAG51 protein.The phase contrast microscope was used on 84 h after transfection to observe the change of the morphology of cultured PC12;Using Hoechst 33258 for staining the nucleus and then observe the nuclear morphological change;the change the morphology of nucleolus was observed with Hoechst33258 coloration;the apoptosis implementing protein,Caspase-3,was examined.Results TDAG51 was overexpressed in PC12;compared with the cells in the two comparison groups,the bodies of the cells with TDAG51 overexpression become round and the neurite was lost;Under fluorescene microscope,the nucleolus of PC12 condensed,the chromatin aggregated and nuclear fragmentation could be seen;the pro-caspase-3 protein was activated incised and,the activated 17 kDa Caspase-3 increased more along with the functioning prolongatisn of TDAG51 protein.Conclusion Overexpression of TDAG51 gene can induce the apoptosis of PC12.

关 键 词：TDAG51 凋亡 PC12 CASPASE-3 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]