CPEB4对慢性髓系白血病细胞迁移与周期的影响

Effect of CPEB4 on Migration and Cycle of Chronic Myeloid Leukemia Cell

目的:探讨CPEB4对慢性髓系白血病细胞K562迁移与周期的影响及可能参与机制调控的蛋白分子的变化。方法:采用Western blot检测正常白细胞和K562细胞中CPEB4的表达水平;再将过表达质粒pcDNA3.1(+)-His-CPEB4、沉默质粒pPLK+Puro-CPEB4 shRNA电穿孔转染K562细胞,改变CPEB4在K562细胞中的表达量,利用Western blot检测转染效率;最后利用Transwell小室、流式细胞术检测不同处理细胞的迁移、周期情况,并用Western blot检测MMP2、MMP9、CDK4、CyclinD1、P21蛋白的表达变化。结果:与正常白细胞相比,K562细胞中的CPEB4蛋白表达量明显升高(P<0.01);CPEB4沉默的K562细胞与对照组相比,细胞的迁移能力显著提高(P<0.01);G0/G1期细胞比例减少,G2/M期细胞比例增加,细胞周期进程加快(P<0.01);MMP2(P<0.05)、MMP9(P<0.05)、CDK4(P<0.01)、CyclinD1蛋白表达水平明显增加(P<0.01),P21蛋白表达显着降低(P<0.01)。CPEB4过表达后K562细胞迁移能力明显下降(P<0.01);细胞周期中G0/G1期细胞比例增加,S期比例降低,细胞周期进程阻滞于G0/G1期(P<0.01);P21蛋白表达明显增加,MMP2、MMP9、CDK4、CyclinD1蛋白表达均显著降低(P<0.05-0.01)。结论:CPEB4可抑制K562细胞迁移,将细胞周期进程阻滞于G0/G1期;CPEB4影响K562细胞周期和迁移可能是通过调控MMP2、MMP9、CDK4、CyclinD1、P21等分子的表达完成的。

Objective:To investigate the effects of CPEB4 on the migration and cycle of K562 cells and the changes of protein molecules that may be involved in the regulatory mechanism.Methods:Western blot was used to detect the expression of CPEB4 in normal leukocytes and K562 cells.The overexpression plasmid pcDNA3.1(+)-His-CPEB4,silencing plasmid pPLK+Puro-CPEB4 shRNA were transfected into K562 cells by electroporation so as to change CPEB4.The transfection efficiency was detected by Western blot.Finally,the migration and cycle of different cells were detected by Transwell chamber and flow cytometry.Western blot was used to detect the expression changes of MMP2,MMP9,CDK4,CyclinD1 and P21 proteins.Results:Compared with normal white blood cells,the expression of CPEB4 protein in K562 cells was significantly enhanced(P<0.01);Compared with the control group,CPEB4-silenced K562 cells showed that the cell migration ability was significantly enhanced(P<0.01);G0/G1 phase cell ratio reduced,G2/M phase cell ratio increased,and cell cycle progression accelerated(P<0.01),The expression levels of MMP2(P<0.05),MMP9(P<0.05),CDK4(P<0.01),CyclinD1(P<0.01)proteins increased significantly.The expression level of P21 protein significantly decreased(P<0.01).The migration ability of K562 cells after CPEB4 overexpression was decreased(P<0.01),the cell ratio of G0/G1 phase in the cell cycle increased,the cell proportion of S phase decreased and the cell cycle progression was arrested at G0/G1 phase(P<0.01).The expression of P21 protein increased,MMP2,MMP9,CDK4,CyclinD1 protein expression decreased significantly(P<0.05-0.01).Conclusion:CPEB4 can inhibit the migration of K562 cells and arrest cell cycle progression at G0/G1 phase.Its mechanism may be related with regulating the exprossion of MMP2,MMP9,CDK4,CyclinD1 and P21 proteins.