WWOX基因对人急性淋巴细胞性白血病细胞CCRF-CEM细胞凋亡及细胞周期的影响

Effect of WWOX on the regulation of the cell cycle and apoptosis in human acute lymphoblastic leukemia cell CCRF-CEM

目的:研究WWOX基因对人急性淋巴细胞性白血病细胞CCRF-CEM细胞凋亡及细胞周期的影响及分子机制。方法:体外培养CCRF-CRM细胞系,将WWOX基因转染入CCRF-CEM细胞,建立高表达WWOX的CCRF-CEM细胞,流式细胞仪法测定CCRF-CEM细胞的细胞周期和凋亡,Western blot法测定WWOX基因、细胞周期和凋亡相关因子的蛋白表达,qRT-PCR法测定caspase-3的mRNA表达。结果:与对照组和空白质粒组相比,转染入WWOX的CCRF-CEM细胞的WWOX蛋白表达显著增加,G_0/G_1期的细胞比例显著增加,凋亡细胞比例显著增加;CCRF-CEM细胞中cyclin D1、cyclin E、CDK2的蛋白表达显著降低而Wnt-5α和JNK的蛋白表达显著增加,caspase-3的mRNA表达促进CCRF-CEM细胞的凋亡。结论:WWOX基因在急性淋巴细胞性白血病CCRF-CEM细胞中发挥抑癌基因作用,可以有效促进细胞凋亡并抑制细胞周期进程。

Objective：To study the effect of WWOX on the regulation of the cell cycle and apoptosis in human acute lymphoblastic leukemia cell CCRF-CEM and the mechanism.Methods：CCRF-CEM cells were transfected with a eukaryotic expression vector carrying WWOX gene.Flow cytometry was used to detect the cell cycle and apoptosis of CCRF-CEM cells.Western blot was used to detect the protein expression of cell regulatory proteins and apoptosis-related proteins.qRT-PCR was ued to detect the mRNA expression of caspase-3.Results：Compared with control group and blank plasmid group，the WWOX protein expression in WWOX-transfected CCRF-CEM cells was increased significantly，and the proportion of cells in G0/G1 phase was increased significantly，and the proportion of apoptotic cells was increased significantly.Cyclin D1，cyclin E，CDK2 protein expression was significantly reduced,but Wnt-5α and JNK protein expression was significantly increased.Caspase-3 mRNA expression promoted CCRF-CEM cell apoptosis.Conclusion：WWOX gene plays a role of tumor suppressor gene in acute lymphoblastic leukemia CCRF-CEM cells，which can effectively promote apoptosis and inhibit cell cycle progression.