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目的:探讨珊瑚树vibsane型二萜类化合物对肝癌HepG2细胞增殖的影响及其机制,为研发新型天然植物类抗肿瘤药物提供实验依据。方法:采用噻唑蓝比色法及苔盼蓝染色计数法观察珊瑚树vibsane类二萜类化合物对不同肿瘤细胞增殖的影响;应用流式细胞仪检测细胞周期及细胞凋亡,利用Apo-ONE Homogeneous Caspase-3/7试剂盒检测vibsane二萜类化合物1#对HepG2细胞内Caspase-3酶活性的影响。结果:活性筛选发现vibsane型二萜类化合物1#显著抑制人肝癌HepG2细胞增殖,构效分析表明化合物C11位连接侧链的基团修饰影响其细胞增殖抑制活性。此外,HepG2细胞对1#化合物最敏感,1#化合物抑制其增殖具有剂量和时间依赖性。机制研究显示1#化合物诱导HepG2细胞发生明显的细胞周期G0/G1期阻滞,具有时间和剂量效应;同时,较高浓度1#化合物(5~10μmol/L)引起HepG2细胞凋亡明显增加,并剂量依赖性诱导细胞内Caspase3/7激活。结论:珊瑚树vibsane型二萜类化合物能够明显抑制人肝癌HepG2细胞增殖,其可能通过诱导细胞周期阻滞和细胞凋亡发挥抗肿瘤作用。
OBJECTIVE: To investigate the effect and mechanism of vibrisane-type diterpenes on proliferation of hepatocellular carcinoma HepG2 cells and to provide experimental evidence for the development of novel natural plant antitumor drugs. Methods: Thiazolyl blue colorimetric assay and trypan blue staining method were used to observe the effects of vibrisane diterpenoids on the proliferation of different tumor cells. Flow cytometry was used to detect the cell cycle and apoptosis. Apo-ONE Homogeneous The Caspase-3/7 kit was used to detect the effect of vibsane diterpenoid 1 # on the activity of Caspase-3 in HepG2 cells. Results: The result of active screening showed that vibsane diterpenoid 1 # significantly inhibited the proliferation of HepG2 cells. The structure-activity analysis indicated that the modification of the side chain of compound C11 affected the cell proliferation inhibitory activity. In addition, HepG2 cells are most sensitive to 1 # compounds, and 1 # compounds inhibit their proliferation in a dose- and time-dependent manner. Mechanistic studies showed that the 1 # compound induced obvious arrest of G0 / G1 phase in HepG2 cells with time and dose effect. At the same time, higher concentration of 1 # compound (5 ~ 10μmol / L) induced a significant increase of apoptosis in HepG2 cells, And dose-dependently induce intracellular Caspase 3/7 activation. CONCLUSION: The coral tree vibsane diterpenoids can significantly inhibit the proliferation of HepG2 cells, which may play an anti-tumor role by inducing cell cycle arrest and apoptosis.