AICAR inhibits proliferation and induced S-phase arrest,and promotes apoptosis in CaSki cells  

作　　者：Tong-ju GUAN Feng-jin QIN Jian-hai DU Li Geng You-yi ZHANG Min LI 

机构地区：[1]Institute of Pathology, The School of Basic Medicine, Lanzhou University, Lanzhou 730000, China [2]Department of Gynaecology andObstetrics, Peking University Third Hospital, Beijing 100083, China [3]Institute of Vascular Medicine, Peking University Third Hospital andKey Laboratory of Molecular Cardiovascular Sciences Ministry of Education, Beijing 100083, China

出　　处：《Acta Pharmacologica Sinica》2007年第12期1984-1990,共7页中国药理学报（英文版）

基　　金：This work was supported by the National Key Basic Research Program of China(№ 2006CB503806);the Natural Science Foundation of China(№ 30672466 and C03030401)

摘　　要：Aim： The aim of the present study was to determine the effect of 5-aminoimidazole- 4-carboxamide-ribonucleoside （A/CAR） on proliferation, cell cycle, and apoptosis in the human epithelial cervical cancer cell line CaSki cells. Methods： Cell count and 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay were used to determine cell proliferation and viability. Hoechst 33258 staining was con- ducted to distinguish the apoptotic cells. Cell cycle and Annexin-V/propidium iodide staining were analyzed by fluorescence-activated cell sorting （FACS）. A Western blot assay was used to evaluate the expression of AKT （also known as protein kinase B）, mammalian target of rapamycin （mTOR）, p53, and extracellular signal-regulated kinase （ERK）. Results： A/CAR （500 pmol/L） significantly inhibi- ted the proliferation of CaSki cells treated for 24, 48, and 72 h as determined by cell count. The cells at the Gl and G2 phases were dramatically decreased while cells at the S phase were increased in response to A/CAR treatment for 24, 48, and 72 h, The MTF assay showed less viable cells and Hoechst fluorescent staining showed more apoptotic cells upon AICAR stimulation. The results of the Annexin-V staining demonstrated a time-dependent increase of apoptosis in cells treated with A/CAR for 24, 36, and 48 h. Furthermore, AICAR activated caspase-3 in a time-dependent manner. It was also found that AICAR inhibited the phosphory- lation of AKT and mTOR, which are important kinases regulating cell growth and survival. AICAR stimulation obviously increased the expression of the tumor suppressor p53 and the phosphorylation of ERK. Conclusion： A/CAR inhibited proliferation and induced S phase arrest and promoted apoptosis in CaSki cells, which might be mediated by the dowrtregulation of the AKT/mTOR pathway and the upregulation of the p53/ERK pathway.Aim： The aim of the present study was to determine the effect of 5-aminoimidazole- 4-carboxamide-ribonucleoside （A/CAR） on proliferation, cell cycle, and apoptosis in the human epithelial cervical cancer cell line CaSki cells. Methods： Cell count and 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay were used to determine cell proliferation and viability. Hoechst 33258 staining was con- ducted to distinguish the apoptotic cells. Cell cycle and Annexin-V/propidium iodide staining were analyzed by fluorescence-activated cell sorting （FACS）. A Western blot assay was used to evaluate the expression of AKT （also known as protein kinase B）, mammalian target of rapamycin （mTOR）, p53, and extracellular signal-regulated kinase （ERK）. Results： A/CAR （500 pmol/L） significantly inhibi- ted the proliferation of CaSki cells treated for 24, 48, and 72 h as determined by cell count. The cells at the Gl and G2 phases were dramatically decreased while cells at the S phase were increased in response to A/CAR treatment for 24, 48, and 72 h, The MTF assay showed less viable cells and Hoechst fluorescent staining showed more apoptotic cells upon AICAR stimulation. The results of the Annexin-V staining demonstrated a time-dependent increase of apoptosis in cells treated with A/CAR for 24, 36, and 48 h. Furthermore, AICAR activated caspase-3 in a time-dependent manner. It was also found that AICAR inhibited the phosphory- lation of AKT and mTOR, which are important kinases regulating cell growth and survival. AICAR stimulation obviously increased the expression of the tumor suppressor p53 and the phosphorylation of ERK. Conclusion： A/CAR inhibited proliferation and induced S phase arrest and promoted apoptosis in CaSki cells, which might be mediated by the dowrtregulation of the AKT/mTOR pathway and the upregulation of the p53/ERK pathway.

关 键 词：AICAR PROLIFERATION apoptosis CaSki cells 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]