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

摘要

Aims To study the effects of diterpenoid pekinenal of Euphorbia pekinensis Rupr. on cell prolif-eration, cell cycle phase and apoptosis of hepatoma SMMC-7721 cells and to probe into its anti-cancer mechanisms. Methods MTT assay was used to inves-tigate 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 concentra-tion, apoptotic rate of SMMC-7721 cells increased sig-nificantly, compared with control group, the difference has significant statistical significance ( P < 0. 01 ) . TUNEL method test results showed that different con-centrations of pekinenal in SMMC-7721 cells could in-duce liver cancer cell apoptotic and apoptotic index ( AI) increased significantly ( P <0. 01 ) with the in-crease of drug concentration. Compared with control group, electron microscope found that after the treat-ment, hepatocyte mitochondrial swelling, endoplasmic reticulum expansion, cytoplasm inclusion body forma-tion, part of the nucleus apoptosis, and the more obvi-ous apoptosis appeared with the increase of drug con-centration. Flow cytometry analysis showed that differ-ent concentrations of pekinenal could all make the cell block in S phase. Conclusion Pekinenal has an obvi-ously inhibitory effect on the human liver cancer cells, and there is significant concentration dependence; Pe-kinenal 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 inhibi-tion by inducing liver cancer cells apoptosis, etc.%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。