摘要
目的:研究多西他赛(docetaxel)体外抗肝癌作用及其与氧化还原状态的关系。方法:用不同浓度的多西他赛处理肝癌细胞株(SMMC-7721细胞),用集落形成试验检测多西他赛对SMMC-7721细胞生长抑制作用,通过流式细胞术测定多西他赛对SMMC-7721细胞株细胞周期及凋亡的影响。用DCF/DA作为活性氧(reactive oxygen species,ROS)捕获剂检测用药后细胞内ROS产生。用试剂盒测定细胞谷胱甘肽(glutathione,GSH)水平。结果:多西他赛能抑制SMMC-7721细胞增殖,并呈浓度依赖性;诱导SMMC-7721细胞凋亡和G2/M期阻滞;导致SMMC-7721细胞内ROS产生增多和GSH降低。结论:多西他赛能通过诱导细胞凋亡和G2/M期阻滞而抑制SMMC-7721细胞增殖,用药后细胞内ROS产生增多和GSH降低可能在多西他赛体外抑制SMMC-7721细胞增殖和促进细胞凋亡中起重要作用。
OBJECTIVE To investigate the in vitro anti- HCC（SMMC-7721 cells）activity of docetaxel and its relationship with redox. METHODS The SMMC-7721 cells growth was measured by colony forming test. Cell cycle and apoptosis were analyzed by flow cytometry. After exponential growth phase cells were treated with different doses of docetaxel for 24hrs, incubated with 2＇ ,7＇-dichlo- rofluorescin diacetate （DCF/DA） to estimate the reactive oxygen species（ROS）level. Also the docetaxel treated cells were harvested by trypsinization, counted under phase contrast microscope The ceils suspension was centrifuged and the cell pellets were added with distilled water and thiosalicylic acid to precipitate the protein. After centrifngation the supernatant was used for glutathione（GSH）assay. The control group was not given with docetaxel. RFSULTS Docetaxel inhibited the HCC; cell growth in concentration dependent manners with IC50 （5×10^10 M）. Marked cell apoptosis and G2/M phase arrest were observed alter treatment with docetaxel≥10^-8 M. The SMMC-7721 cells ROS level （corresponding to the fluorescence intensity） increased after treatment with different doses of docetaxel for 24 h. The intracellular GSH content after treatment with different doses of docetaxel for 24 hrs decreased, there was a significant difference between the drug treated groups and the control group （P〈0. 05）. CONCLUSION Docetaxel suppresses the growth of SMMC-7721 human HCC; cells in vitro by causing apoptosis and cell-cycle arrest of the human HCC cells, and the ROS and GSH may play a key role in the inhibition of growth and induction of apoptosis.
出处
《中国医院药学杂志》
CAS
CSCD
北大核心
2008年第6期452-455,共4页
Chinese Journal of Hospital Pharmacy
