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目的 探讨二烯丙基二硫 (DADS)诱导HL 6 0细胞G2 /M期细胞生长阻滞的分子机制。方法 用不同浓度的DADS处理HL 6 0细胞 ,分别作用 0 ,6 ,1 2 ,2 4 ,4 8h后用MTT法测定细胞增殖活性 ;用流式细胞术和有丝分裂指数测定细胞增殖周期 ;用Westernblot检测 p38丝裂原活化蛋白 (p38MAPK)及Cdc2 5B和Cdc2激酶的表达和磷酸化水平 ;用RT PCR检测p38MAPKmRNA的表达。结果 DADS抑制HL 6 0细胞增殖呈浓度依赖性 ,且DADS(2 0 μmol/L)与ATRA(1 0nmol/L)对HL 6 0细胞增殖活性影响相近 (P >0 .0 5 ) ;DADS 2 0 μmol/L作用HL 6 0细胞 1 2h后能引起G2 /M期细胞百分数增高并达到最大值 ,而此时有丝分裂指数明显下降 (P <0 .0 5 ) ;同时 ,磷酸化p38MAPK蛋白及p38MAPKmRNA的表达达到高峰 (P <0 .0 5 ) ,并引起Cdc2 5B和Cdc2磷酸化水平的相应变化。而p38特异性抑制剂SB2 0 2 1 90 (1 0 μmol/L)能阻断DADS对HL 6 0细胞增殖的抑制作用 (P <0 .0 5 )。结论 DADS能启动HL 6 0细胞G2 /M控制点 ,它的激活可能与磷酸化 p38MAPK的活化有关
Objective To investigate the molecular mechanism of diallyl disulfide (DADS) -induced G2 / M cell cycle arrest in HL-60 cells. Methods HL-60 cells were treated with different concentrations of DADS and the cell proliferation activity was determined by MTT assay at 0, 6, 1 2, 2 4 and 4 8 hours respectively. The cell cycle was measured by flow cytometry and mitosis index. Western Blot The expression and phosphorylation of p38 mitogen-activated protein (p38MAPK) and Cdc2 5B and Cdc2 kinase were detected. The expression of p38 MAPK mRNA was detected by RT-PCR. Results DADS inhibited the proliferation of HL-60 cells in a concentration-dependent manner. DADS (20 μmol / L) and ATRA (10 nmol / L) had similar effects on the proliferation of HL-60 cells (P> 0.05) The percentage of G2 / M phase cells was increased and reached its maximum at 0 μmol / L HL 60 cells for 1 h and the mitotic index decreased significantly at this time (P <0.05). Meanwhile, phosphorylated p38MAPK protein and p38MAPK mRNA (P <0.05), and caused the corresponding changes in phosphorylation levels of Cdc2 5B and Cdc2. However, p38-specific inhibitor SB2021 90 (10 μmol / L) blocked the inhibitory effect of DADS on the proliferation of HL-60 cells (P <0.05). Conclusion DADS can activate the G2 / M control point of HL-60 cells, and its activation may be related to the activation of phosphorylated p38MAPK