谷胱甘肽耗竭促成紫花牡荆素诱导肝癌细胞凋亡  

Glutathione depletion con'tributes to casticin-induced apoptosis in hepatocellular carcinoma cells

作　　者：刘跃银 杨军[2] 盛习锋[3] 

机构地区：[1]郴州市第二人民医院,湖南郴州423000 [2]中南大学湘雅三医院,湖南长沙410013 [3]湖南师范大学医学院,湖南长沙410013

出　　处：《肿瘤药学》2011年第4期351-355,共5页Anti-Tumor Pharmacy

基　　金：湖南省中医药管理局科技项目(NO.2010081);湖南省教育厅科技项目(NO.10C0975);湖南省教育厅科技重点项目(NO.09A054)

摘　　要：目的研究紫花牡荆素(casticin)诱导肝癌细胞(HCC)凋亡作用及其作用分子机制。方法体外培养人肝癌PLC/PRF/5细胞系细胞,MTT测定细胞活性;细胞凋亡ELISA检测试剂盒和碘化丙啶(PI)染色流式细胞术(FCM)检测凋亡性细胞死亡。二氯二氢荧光素二乙酯(DCFH-DA)探针标记FCM测定活性氧(ROS)生成。谷胱甘肽检测试剂盒测定细胞内谷胱甘肽(GSH)含量。结果 Casticin以剂量依赖方式抑制PLC/PRF/5细胞生长(P<0.05)。Casticin诱导PLC/PRF/5细胞系Sub-G1细胞百分率增加(P<0.05);并增加细胞组蛋白/DNA碎片的含量,呈浓度依赖性。Casticin降低PLC/PRF/5细胞内GSH含量(P<0.05),但不影响胞内ROS的生成。巯基抗氧化剂,N乙酰半胱氨酸和添加外源性GSH能恢复GSH含量,并减弱casticin诱导细胞凋亡作用;而不含巯基的抗氧化剂,丁羟茴醚和甘露醇无明显作用。结论 Casticin诱导肝癌细胞凋亡涉及细胞内GSH耗竭。Objective To investigate the apoptotic activities of casticin in hepatocellular carcinoma （HCC） cells and its molecular mechanisms.Method Hepatocellular carcinoma PLC/PRF/5 cells were cultured in vitro and the inhibitory effect of casticin on growth of cells was detected by MTT assay.The apoptotic cell death was examined using the cell apoptosis ELISA detection kit and flow cytometry （FCM） after propidium iodide （PI） staining.Reactive oxygen species （ROS） production was evaluated by FCM after dichlorodihydrofluorescein diacetate （DCFH-DA） probe labeling.Intracellular glutathione （GSH） content was measured using a Glutathione Assay kit.Results Casticin significantly inhibited the growth of PLC/PRF/5 cells in a dose-dependent manner （P〈0.05）.Casticin increased the percentage of the sub-G1 population and increased the levels of Histone/DNA fragmentation in PLC/PRF/5 cells,in a concentrationdependent manner （P〈0.05）.Casticin decreased intracellular GSH content （P〈0.05）,but not affected the level of intracellular ROS in PLC/PRF/5 cells.The thiol antioxidants,acetylcysteine and ectogenic GSH restored GSH content and attenuated casticin-induced apoptosis.In contrast,the nonthiol antioxidants,butylated hydroxyanisole and mannitol failed to do so.Conclusion Casticin induced apoptosis of HCC cells is involved in GSH depletion

关 键 词：肝癌 紫花牡荆素 谷胱甘肽 细胞凋亡 

分 类 号：R737.9[医药卫生—肿瘤]