PUMA基因对HELA的作用及其机制的初步研究

To Preliminary Study the Effect and Mechanism of PUMA Gene to HELA Cell

目的初步探讨PUMA基因对HELA细胞是否具有促凋亡的活性和其参与HELA细胞凋亡的过程线粒体中cyt-c、AIF位置是否发生迁移。方法倒置相差显微镜、AO/EB染色检测PUMA基因的促凋亡活性;Westernblot检测HELA细胞凋亡后线粒体中cyt-c、AIF位置是否发生迁移。结果①倒置相差显微镜和荧光显微镜结果显示转染PUMA基因后HELA细胞出现了一系列凋亡形态学改变,这种变化在转染48h比24h更明显;②Western blot结果显示HELA细胞凋亡后线粒体内膜腔隙蛋白cyt-c、AIF向胞质迁移,迁移的蛋白量在转染48h比24h更明显。结论①PUMA基因有促进HELA细胞凋亡的作用;②AIF和cyt-c参与PUMA基因促进HELA细胞凋亡的过程,但此过程有无涉及线粒体膜电位的变化还有待进一步的研究。

Objective To preliminary investigate the cytoactive of HELA cell after PUMA gene promoting HELA cell apoptosis and whether or not the protein of cyt - c, AIF in the chondrosome have participated in the process of PUMA gene inducing HELA cell. Methods we Used AO/EB dyeing to detect the Morphologic change of cell induces by PUMA gene and Western Blot to detect whether or not the position of cyt - c,AIF in the chondrosome have transferred after PUMA gene induces HELA cell apoptosis. Result （1）Fluorescence mi- croscope and inverted phase contrast microscope display HELA cell have a series of apoptosis Morphologic change after transfecting PUMA into HELA cell,and this Morphologic change of cell is more obviously after transfecting 48h than after 24h. ;（2）Western Blot display the protein cyt - c and AIF transferred toward to kytoplasm after HELE cell took place apoptosis, the mount of transfecting protein is more obviously after transfecting 48h than after 24h. Conclusion （1） PUMA gene have the effect of promoting HELA cell apoptosis;（2）cyt - c and AIF take part in the process of promoting HELA cell apoptosis, but during this process if the membrane potential of chondrosome was chan- ging need to do some more study to confirm it.