姜黄素对Hep 1-6肝癌细胞凋亡及ROS影响（.docVIP

姜黄素对Hepa 1-6肝癌细胞凋亡及ROS影响( 杜琴，邓珊，沈克平，胡兵(( （上海中医药大学附属龙华医院肿瘤科，中医肿瘤研究所，上海 20032） 摘 要 目的：观察姜黄素对小鼠肝癌Hepa 1-6细胞凋亡及活性氧(Reactive oxygen species， ROS)水平的影响。方法：不同浓度姜黄素处理Hepa 1-6肝癌细胞， MTT比色法检测细胞增殖，Hoechst 33258染色检测细胞凋亡形态变化，流式细胞术检测细胞凋亡，Western Blot法检测活化型caspase-3， 2-7-二氯氢化荧光素乙二脂（DCFH-DA）染色法结合荧光酶标仪检测姜黄素对胞内ROS产生的影响利用荧光探针DCFH-DA进行活性氧检测利用荧光探针DCFH-DA进行活性氧检测利用荧光探针DCFH-DA进行活性氧检测。结果：2～40 μmol/L姜黄素作用24 h、48 h均可抑制肝癌Hepa 1-6细胞增殖，并呈现一定的时间与剂量依赖性。5～20 μmol/L姜黄素作用48 h后Hepa 1-6细胞呈现凋亡形态变化，细胞凋亡率呈一定程度剂量依赖性，同时可检测到活化型caspase-3、以及ROS产生。结论：姜黄素可以抑制Hepa 1-6细胞增殖，促使Hepa 1-6细胞凋亡，可能与活化caspase-3及提高细胞内ROS水平相关。 关键词 肝癌；姜黄素；细胞凋亡；caspase-3；ROS Effects of Curcumin on ROS production and apoptosis in Murine Hepatocarcinoma Hepa 1-6 Cells Du Qin, Deng Shan, Shen Keping, Hu Bing （Department of Oncology and Institute of Traditional Chinese Medicine in Oncology, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, 20032）Objective：To observe the effects of curcumin on apoptosis and reactive oxygen species (ROS) production in murine hepatocarcinoma Hepa1-6 cells in vitro. Methods：Hepa1-6 cells were treated with different dose of curcumin. Cell proliferation was detected with MTT assay. Apoptotic morphology was visualized by hoechst 33258 staining. Cell apoptosis was detected by flow cytometry analysis. Caspase-3 was detected by western blot. Intracellular ROS production was detected by 2’, 7’-dichlorofluorescin diacetate (DCFH-DA) staining. Results：Compared with the controls, upon treatment with 2~40 μmol/L of curcumin for 24 h to 48 h, Hepa 1-6 cells proliferation was significantly inhibited in a time and dose dependent manner. 5~20 μmol/L of curcumin also induced apoptosis and apoptotic morphology change in Hepa 1-6 cells. After treatment with 5~20 μmol/L curcumin, caspase-3 was activated accompanied by generation of ROS. Conclusion：Curcumin may inhibit cell proliferation, induce apoptosis in murine hepatocarcinoma Hepa1-6 cells, and may involvement of caspase-3 activation and ROS prod