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目的:探讨岩大戟内酯B(jolkinolide B,JB)对人乳腺癌MDA-MB-231细胞增殖及生长周期的影响。方法:取对数生长期的MDA-MB-231细胞,加入JB,使终质量浓度分别为0,2.5,5,10,20,40,80 mg·L-1,采用四甲基偶氮唑蓝(MTT)检测岩大戟内酯B对MDA-MB-231细胞的生长抑制率。另设10,20,40,80 mg·L-1组的JB,采用流式检测技术分析岩大戟内酯B对MDA-MB-231细胞的细胞周期,RT-PCR及Western blot检测真核细胞翻译起始因子(e IF4E),细胞周期蛋白D1(Cyclin D1)基因mRNA表达及蛋白水平。结果:随着岩大戟内酯B浓度增加可显著抑制MDA-MB-231细胞的增殖,G1期细胞阻滞。与空白组比较,20,40μmol·L-1的岩大戟内酯B可抑制Cyclin D1和e IF4E的mRNA表达,转染e IF4E-siRNA组Cyclin D1的mRNA表达及蛋白水平也被抑制,同时细胞周期阻滞在G1期(P<0.05,P<0.01)。结论:岩大戟内酯B可通过下调e IF4E及Cyclin D1的表达而阻滞细胞周期于G1期。
Objective: To investigate the effect of jolkinolide B (JB) on proliferation and cell cycle of human breast cancer MDA-MB-231 cells. Methods: The logarithmic growth phase MDA-MB-231 cells were treated with JB to make the final concentrations of 0, 2.5, 5, 10, 20, 40 and 80 mg · L-1, respectively. Blue (MTT) was used to detect the inhibitory effect of petrofurm B on MDA-MB-231 cells. Another group of 10, 20, 40, 80 mg · L-1 groups of JB, using flow cytometry analysis of esmolol B MDA-MB-231 cell cycle, RT-PCR and Western blot detection of eukaryotic Cell translation initiation factor (e IF4E), cyclin D1 gene mRNA expression and protein levels. Results: The proliferation of MDA-MB-231 cells was significantly inhibited as the concentration of petrofurm B increased, and the cell cycle arrest in G1 phase. Compared with the blank group, CODC1 inhibited the mRNA expression of Cyclin D1 and e IF4E at 20 and 40 μmol·L-1, and inhibited the mRNA and protein expression of Cyclin D1 in eIF4E-siRNA transfected group Cell cycle arrest in G1 phase (P <0.05, P <0.01). CONCLUSION: Inhibolide B can block the cell cycle in G1 phase by down-regulating eIF4E and Cyclin D1 expression.