京大戟中二萜类化合物pekinenal对肝癌细胞增殖、周期和凋亡的影响  

Effects of diterpenoid pekinenal of Euphorbia pekinensis Rupr.on proliferation,cell cycle and apoptosis of hepatoma cells

作　　者：陈飞燕[1] 陶伟伟[1,2,3] 陈扣玉 张嘉卿[2] 王雨青[2] 陈刚[2] 段金廒[3] 

机构地区：[1]南京中医药大学基础医学院实验研究中心,江苏南京210023 [2]南京中医药大学中医脑病重点实验室,江苏南京210023 [3]南京中医药大学江苏省中药资源产业化过程协同创新中心,江苏南京210023

出　　处：《中国药理学通报》2016年第4期519-524,共6页Chinese Pharmacological Bulletin

基　　金：国家重点基础研究发展计划(973计划)资助项目(No2011CB505303);国家自然科学基金资助项目(No81403041;81274199);江苏省自然科学基金青年基金(NoBK20140961)

摘　　要：目的研究京大戟中二萜类化合物pekinenal对肝癌SMMC-7721细胞增殖、周期和凋亡的影响,并初步探讨其治疗肝癌可能的影响机制。方法采用MTT法分析不同浓度的pekinenal对体外培养SMMC-7721细胞增殖的影响;采用原位末端标记法(TUNEL)、Annexin V/PI双染法和电镜观察对细胞凋亡的作用;应用流式细胞术分析对细胞周期的影响。结果 MTT结果显示,pekinenal可明显抑制SMMC-7721细胞的增殖,呈浓度依赖性。Annexin V/PI双染法结果显示,随着pekinenal浓度的增加,肝癌SMMC-7721细胞的凋亡率明显增加,与对照组比较,差异具有统计学意义(P<0.01)。TUNEL法检测结果表明,不同浓度的pekinenal作用于SMMC-7721细胞后均可诱导肝癌细胞凋亡,凋亡指数(AI)随着药物浓度的升高明显增加(P<0.01)。给药后电镜观察,与对照组比较,发现肝细胞线粒体肿胀、内质网扩张,胞质包涵体形成,部分细胞核呈现凋亡表现,并且随着药物浓度增加,凋亡现象越明显。流式细胞术分析显示,不同浓度的pekinenal均可使细胞被阻滞于S期。结论 pekinenal对人肝癌细胞增殖有明显地抑制作用,并存在着明显的浓度依赖关系;pekinenal可能是通过抑制癌细胞DNA的合成,将人肝癌细胞周期阻滞在S期,抑制其增殖,通过诱导人肝癌细胞发生凋亡等,发挥其抗肝癌作用。Aims To study the effects of diterpenoid pekinenal of Euphorbia pekinensis Rupr. on cell proliferation,cell cycle phase and apoptosis of hepatoma SMMC-7721 cells and to probe into its anti-cancer mechanisms. Methods MTT assay was used to investigate the effect of pekinenal on proliferation of SMMC-7721 cells; TUNEL method,Annexin V / PI staining and electron microscopy were employed to observe the cell apoptosis; Flow cytometry was applied to analyze the distribution of cell cycle. Results Proliferation of SMMC-7721 cells was markedly inhibited by pekinenal in a dose-dependent manner; Annexin V / PI staining showed that with the increase of pekinenal concentration,apoptotic rate of SMMC-7721 cells increased significantly,compared with control group,the difference has significant statistical significance( P < 0. 01).TUNEL method test results showed that different concentrations of pekinenal in SMMC-7721 cells could induce liver cancer cell apoptotic and apoptotic index( AI) increased significantly( P < 0. 01) with the increase of drug concentration. Compared with control group,electron microscope found that after the treatment,hepatocyte mitochondrial swelling,endoplasmic reticulum expansion,cytoplasm inclusion body formation,part of the nucleus apoptosis,and the more obvious apoptosis appeared with the increase of drug concentration. Flow cytometry analysis showed that different concentrations of pekinenal could all make the cell block in S phase. Conclusion Pekinenal has an obviously inhibitory effect on the human liver cancer cells,and there is significant concentration dependence; Pekinenal probably inhibits cancer cell DNA synthesis for the human liver cell cycle arrest in S phase and inhibits the proliferation. It plays a role in liver cancer inhibition by inducing liver cancer cells apoptosis,etc.

关 键 词：京大戟 pekinenal SMMC-7721 细胞增殖 细胞周期 细胞凋亡 

分 类 号：R284.1[医药卫生—中药学] R329.24[医药卫生—中医学]