Nudix水解酶21对白血病K562细胞增殖的影响

Effect of Nudix hydrolase 21 on proliferation of leukemia K562 cells

目的研究m RNA前体3′-端修饰因子Nudix水解酶21(Nudt21)对白血病K562细胞增殖的影响。方法通过RT-PCR和蛋白质印迹法检测白血病细胞系中Nudt21 m RNA和蛋白表达水平;慢病毒介导RNA干扰下调K562细胞Nudt21表达,通过CCK8检测细胞增殖活力、流式细胞仪检测细胞周期和凋亡水平、蛋白质印迹法检测细胞增殖、凋亡及细胞周期相关蛋白表达。结果在白血病细胞K562、Jurkat和HL-60中Nudt21 m RNA表达高于正常对照组(P<0.01);Nudt21蛋白水平高于正常对照组(P<0.01)。通过慢病毒介导的Nudt21干扰后,K562细胞CCK8掺入量降低(P<0.01)。Nudt21干扰组细胞凋亡率增高(P<0.01)。Nudt21干扰组G0/G1期的细胞增多(P<0.01),S期的细胞数降低(P<0.01)。Ki67蛋白表达下降(P<0.01)。凋亡促进蛋白Bax表达升高(P<0.01)。凋亡抑制蛋白Bcl-2表达下降(P<0.01)。细胞周期蛋白E表达下降。结论 Nudt21可促进K562细胞增殖、抑制凋亡,影响细胞周期分布。

Objective To investigate the effect of Nudt21, a 3′-end-modifier of precursor mRNA, on the proliferation of leukemia K562 cells. Methods The expression of Nudt21 mRNA and protein in several leukemia cell lines was detected by RT-PCR and Western blotting. Then, the expression of Nudt21 in K562 cells was down-regulated with RNA interference mediated bylentivirus. Cell viability was detected by CCK8, cell cycle and apoptosis by flow cytometry, and cell proliferation, apoptosis, and cell cycle-related protein expression by Western blotting. Results The expression of Nudt21 mRNA in leukemia cell lines K562, Jurkat and HL-60 was higher than that in the normal control group（P〈0.01）, and so was the level of Nudt21 protein（P〈0.01）. The incorporation of CCK8 in K562 cells was reduced by lentiviral-mediated Nudt21 interference（P〈0.01）. The Nudt21 interference group showed higher apoptosis rate（P〈0.01）, more cells in the G0/G1 phase（P〈0.01）, fewer cells in the S phase（P〈0.01）, less Ki67 protein expression（P〈0.01）, enhanced expression of apoptosis-promoting protein Bax（P〈0.01）, less expression of apoptosis-inhibitory protein Bcl-2（P〈0.01） and cyclin E. Conclusion Nudt21 may interfere with cell cycle distribution of K562 cells by promoting cell proliferation and inhibiting apoptosis.