Punicalagin induces apoptotic and autophagic ce death in human U87MG glioma cells  

作　　者：Shyang-guang WANG Ming-hung HUANG Jui-hsiang LI Fu-i LAI Horng-mo LEE Yuan-nian HSU 

机构地区：[1]Institute of Pharmaceutical Science and TechnoloEy, Central Taiwan University of Science and Technology, TaichunE,, Taiwan, China [2]Department of Medical Laboratory Science and Biotechnology, School of Medicine, Taipei Medical University, Taipei, Taiwan, China [3]Department of Health, Taoyuan General Hospital, Taoyuan, Taiwan, China [4]Department of Medical Laboratory Science and Biotechnology, Central Taiwan University of Science and Technology, Taichung, Taiwan, China

出　　处：《Acta Pharmacologica Sinica》2013年第11期1411-1419,共9页中国药理学报（英文版）

摘　　要：Aim： To investigate the effects of punicalagin, a polyphenol isolated from Punica granatum, on human U87MG glioma cells in vitro. Methods： The viability of human U87MG glioma cells was evaluated using MTT assay. Cell cycle was detected with flow cytometry analysis. The levels of Bcl-2, cleaved caspase-9, cleaved poly（ADP-ribose） polymerase （PARP）, phosphor-AMPK and phosphor-p27 at Thr198 were measured using immunoblot analyses. Caspase-3 activity was determined with spectrophotometer. To determine autopha^Lv, LC3 cleavage and punctate patterns were examined. Results： Punicalagin （1-30 pp=VmL） dose-dependently inhibited the cell viability in association with increased cyclin E level and decreased cyclin B and cyclin A levels. The treatment also induced apoptosis as shown by the cleavage of PARP, activation of caspase-9, and increase of caspase-3 activity in the cells. However, pretreatment of the cells with the pan-caspase inhibitor z-DEVD- fmk （50 pmol/L） did not completely prevent the cell death. On the other hand, punicalagin treatment increased LC3-11 cleavage and caused GFP-LC3-11-stained punctate pattern in the cells. Suppressing autopha~, of cells with chloroquine （1-10 pmol/L） dose- dependently alleviated the cell death caused by punicalagin. Punicalagin （1-30 pp=VmL） also increased the levels phosphor-AMPK and phosphor-p27 at Thr198 in the cells, which were correlated with the induction of autophagic cell death. Conclusion： Punicalagin induces human U87MG glioma cell death through both apoptotic and autophagic pathways.Aim： To investigate the effects of punicalagin, a polyphenol isolated from Punica granatum, on human U87MG glioma cells in vitro. Methods： The viability of human U87MG glioma cells was evaluated using MTT assay. Cell cycle was detected with flow cytometry analysis. The levels of Bcl-2, cleaved caspase-9, cleaved poly（ADP-ribose） polymerase （PARP）, phosphor-AMPK and phosphor-p27 at Thr198 were measured using immunoblot analyses. Caspase-3 activity was determined with spectrophotometer. To determine autopha^Lv, LC3 cleavage and punctate patterns were examined. Results： Punicalagin （1-30 pp=VmL） dose-dependently inhibited the cell viability in association with increased cyclin E level and decreased cyclin B and cyclin A levels. The treatment also induced apoptosis as shown by the cleavage of PARP, activation of caspase-9, and increase of caspase-3 activity in the cells. However, pretreatment of the cells with the pan-caspase inhibitor z-DEVD- fmk （50 pmol/L） did not completely prevent the cell death. On the other hand, punicalagin treatment increased LC3-11 cleavage and caused GFP-LC3-11-stained punctate pattern in the cells. Suppressing autopha~, of cells with chloroquine （1-10 pmol/L） dose- dependently alleviated the cell death caused by punicalagin. Punicalagin （1-30 pp=VmL） also increased the levels phosphor-AMPK and phosphor-p27 at Thr198 in the cells, which were correlated with the induction of autophagic cell death. Conclusion： Punicalagin induces human U87MG glioma cell death through both apoptotic and autophagic pathways.

关 键 词：PUNICALAGIN human glioma autophagy apoptosis microtubule-associated protein light chain 3 AMPK P27 z-DEVD-fmk CHLOROQUINE 

分 类 号：Q786[生物学—分子生物学] Q251