人类肿瘤细胞分化中细胞周期素的表达规律及作用机制

The expression and effects of Cyclins in inducing human tumor cell differentiation

目的 探讨诱导人类肿瘤细胞分化过程中 ,细胞周期素 (Cyclin)的变化在细胞周期调控机制中的作用。方法 应用全反式维甲酸 (ATRA)处理HL 6 0细胞 ,流式细胞仪行细胞周期分析 ,应用十二烷基磺酸钠 聚丙烯胺凝胶电泳 (SDS PAGE)及WesternBlot检测细胞周期素D1、E的蛋白表达量改变 ,使用流式细胞仪检测细胞周期素D1、E、A的表达量变化及细胞周期素B的时相性改变。结果 HL 6 0细胞经ATRA诱导后 ,80 %左右的细胞被阻滞在G0 G1 期 ,S期细胞减少到2 .81%。WesternBlot显示 ,诱导后 ,CyclinD1表达量明显降低 ,CyclinE表达量升高。流式细胞仪检测CyclinD1、CyclinE蛋白表达变化与WesternBlot结果相同 ,CyclinA表达量升高 ;CyclinB表达特征由经诱导前的非时相性表达转为诱导分化后的时相性表达。结论 细胞周期素D1、E、A、B与肿瘤细胞分化的细胞周期调控有着密切的关系。

Objective To study the changes of Cyclins in all-trans retinoic acid-induced HL-60 cell differentiation and then to investigate the cell cycle regulation in inducing tumor cell differentiation.Methods The effects of all-trans retinoic acid at the dosage of 1×10 -5 mol/L on the differentiation were examined by cytometry analysis.Western Blot was used to observe the changes of Cyclin D1 and Cyclin E in HL-60 cells treated with ATRA.Meanwhile,the amounts of Cyclin D1,cyclin E,Cyclin A,cyclin B expression and the characteristics of cyclin B expression were detected by flow cytometry.Results ATRA inhibited the growth of HL-60 cells.About 80 % cells were arrested at G 0/G 1 phase.The amounts of Cyclin D1 were decreased and the amounts of Cyclin E were increased in ATRA-treated HL-60 cells with Western Blot.The flow cytometic examination of HL-60 cells treated with ATRA revealed that the amounts of Cyclin D1 were decreased,but the amounts of Cyclin E and Cyclin A were increased.Novelly,the characteristics of Cyclin B expression was from unscheduled expression to scheduled one.Conclusion Cyclin D1,Cyclin E,Cyclin A and Cyclin B may be associated with the cell cycle regulation in inducing tumor cell differentiation.