青蒿琥酯对人肺腺癌A549细胞中caspase-9及caspase-3活性的影响  

Apoptosis Induced by Artesunate in Human Adenocarcinoma Cell Line A549 Cells Associated with Activation of caspase-9 and caspase-3

作　　者：王燕燕[1] 宋兴福[1] 崔向军[1] 黄骥[1] 

机构地区：[1]三峡大学第一临床医学院,宜昌市中心人民医院,湖北宜昌443003

出　　处：《肿瘤防治研究》2007年第9期651-653,共3页Cancer Research on Prevention and Treatment

基　　金：湖北省自然科学基金资助项目(2006ABA205)

摘　　要：目的研究青蒿琥酯(artesunate,Art)体外诱导人肺腺癌A549细胞凋亡及对半胱天冬氨酸蛋白酶9(cysteine containing aspartate9,caspase-9)和caspase-3活性的影响。方法Art处理A549细胞,流式细胞计数(Flow Cytometry,FCM)检测细胞周期和细胞凋亡,比色法检测caspase-9活性,westernblot检测caspase-3变化。结果FCM显示A549细胞经100mg/L Art作用24h,出现S期细胞减少(P<0.01),G2/M期细胞数目增多(P<0.01),细胞凋亡率增加(P<0.0)。不同浓度(10mg/L、25mg/L、50mg/L、100mg/L)的Art作用A549细胞24h,caspase-9活性呈浓度依赖性增加,分别为对照组的9.87倍、23.33倍(P<0.01)、38.47倍(P<0.01)和60.47倍(P<0.01)。Western blot显示A549细胞经100mg/L Art作用6、12、24h后,细胞浆中caspase-3活化,分别为对照组1.2倍、1.6倍(P<0.01)、1.8倍(P<0.01)。结论青蒿琥酯可通过增加caspase-9和caspase-3的活性,诱导A549细胞凋亡,为阐明Art的抗癌机理,指导青蒿琥酯用于肿瘤治疗提供实验依据。Objective To investigate the effects of artesunate on apoptosis of human adenocarcinoma cell line A549 cells and activation of cysteine containing aspartate 9（caspase-9） and cysteine containing asparrate 3（caspase-3）. Methods A549 cells were treated with artesunate, cell cycle phases distribution and apoptosis were detected by flow cytometry. Activation of caspase-9 was identified by colorimetric assay. Change of caspase-3 was detected by western blot. Results With artesunate treatment, marked cell accumulation in G2/M phase was observed after 100 mg/L Art exposure for 2411. In addition, the percentage of apoptosis increased, from （3.5 ± 0. 7） % in control cultures to approximately （19.4 ± 1.5） % after 100 mg/L Art treated for 24 h caspase-9 colorimetric assay showed basal levels of caspase-9 activity in A549 cells, and after Art treatment, caspase-9 activity increased in a dose-dependent manner. After A549 cells treated with 100 mg/L Art for 6 h, 12 h and 24 h, the activity of caspase-3 increased with fold of 1.2, 1.6（P〈0.01） and 1.8 （P〈0. 01）, respectively. Conclusion Art is of strong anticancer effect which is associated with activation of caspase-9 and caspase-3 specific pathway.

关 键 词：青蒿琥酯 A549细胞 细胞凋亡 CASPASE-9 CASPASE-3 

分 类 号：R734.2[医药卫生—肿瘤]