摘要
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
Objective To explore the molecular mechanisms of G 2/M checkpoint initiated by diallyl disulfide(DADS) in HL 60 cells. Methods Cell viability was determined by MTT assay. Cell cycle was assayed by flow cytometry. The expression of phospho p38, Cdc25B and Cdc2, and p38 mRNA were measured by Western blotting and RT PCR, respectively. Results After treatment with DADS at 5 ～160 μmol/L for 0～72 h, the growth of HL 60 cells were suppressed in a concentration dependent manner and the inhibitory effect of DADS(20 μmol/L) was similar to that of ATRA(10 nmol/L) (P>0.05). Incubation of HL 60 cells with DADS(20 μmol/L) for 12 h could activate G 2/M checkpoint and increase the expression of phospho p38 MAPK, followed by the expression of phospho Cdc25B and phospho Cdc2(P<0.05). SB202190, a specific inhibitor of p38 MAPK, markedly blocked the phosphorylation of p38 MAPK , Cdc25B and Cdc2(P<0.05). Conclusion DADS could induce the G 2/M arrest in HL 60 cells which may be involved in the activation of p38 MAP kinase.
出处
《中华血液学杂志》
CAS
CSCD
北大核心
2004年第5期273-276,共4页
Chinese Journal of Hematology
基金
国家自然科学基金资助项目 ( 3 0 2 70 684)
湖南省杰出中青年专家专项基金资助项目 ( 0 2JJYB0 0 4)
湖南省自然科学基金资助项目 ( 0 3JJY3 0 2 8)
