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目的研究对映-贝壳杉烷型二萜Wangzaozin A对人早幼粒白血病细胞HL-60生长抑制、周期阻滞、凋亡诱导作用以及相关的机制。方法利用倒置显微镜观察和台盼蓝排染法检测了Wangzaozin A对HL-60细胞生长的影响;采用流式细胞术和吖啶橙/溴化乙锭(AO/EB)双染法检测化合物对细胞周期分布的影响以及诱导HL-60细胞凋亡的能力;进一步利用单细胞凝胶电泳法测试化合物对细胞DNA的损伤。结果 1～4μmol/L的Wangzaozin A对HL-60细胞具有较强的生长抑制作用,较高浓度(3μmol/L和4μmol/L)和较长处理时间(48～72 h)有较强的致死效应;1～2.5μmol/L药物处理细胞24 h或48 h条件下细胞周期分布主要表现为G1期阻滞;4μmol/L药物处理12～60 h条件下细胞周期分布主要表现为很强的S期阻滞;2～4μmol/L药物作用24 h或48 h可诱导凋亡产生,其凋亡率呈显著水平(P<0.01),相同条件下,药物对细胞DNA的损伤与对照相比差异有统计学意义(P<0.05,P<0.01)。结论该化合物对HL-60细胞DNA的损伤作用极可能是引起细胞生长抑制、周期阻滞及凋亡的直接原因。
Aim To study the growth inhibition, cell cycle arrest and apoptosis induction of the human promyelocytic leukemia cell line HL-60 by the ent-kaurene diterpene Wangzaozin A and its related mechanism. Methods The effect of Wangzaozin A on the growth of HL-60 cells was examined by inverted microscope and trypan blue exclusion. The inhibitory effect of Wangzaozin A on the growth of HL-60 cells was detected by flow cytometry and AO / EB staining. Cell cycle distribution and the ability to induce apoptosis of HL-60 cells; further use of single cell gel electrophoresis test compounds for DNA damage. RESULTS: Wangzaozin A (1 ~ 4μmol / L) had a strong inhibitory effect on HL-60 cells. The higher concentrations of 3μmol / L and 4μmol / L and the longer treatment time (48 ~ 72h) The cell cycle distribution of cells treated with 1 ~ 2.5μmol / L of drug for 24 h or 48 h mainly showed the arrest of G1 phase. The cell cycle distribution of cells treated with 4 μmol / L of drug for 12 ~ 60 h mainly showed strong S (P <0.01). Under the same conditions, the DNA damage of the drug was significantly lower than that of the control group There was statistical significance (P <0.05, P <0.01). Conclusion The DNA damage of HL-60 cells is most likely to be the direct cause of cell growth inhibition, cell cycle arrest and apoptosis.