CDK1干扰调节PLK1、Aurora B和TRF1对白血病细胞增殖的影响  

Effect of CDK1 Interferes with the Regulation of PLK1, Aurora B and TRF1 on the Proliferation of Leukemia Cells

作　　者：王冲[1] 李梦亚 申晓辉 王树娟[1] 王伟琼[1] 刘延方[1] WANG Chong;LI Meng-Ya;SHEN Xiao-Hui;WANG Shu-Juan;WANG Wei-Qiong;LIU Yan-Fang(Department of Hematology,The First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,Henan Province,China)

机构地区：[1]郑州大学第一附属医院血液科,河南郑州450052

出　　处：《中国实验血液学杂志》2021年第4期1129-1135,共7页Journal of Experimental Hematology

基　　金：国家自然科学基金项目(U1804191,81800137);河南省科技攻关计划项目(182102310563)。

摘　　要：目的:研究CDK1干扰调节PLK1、Aurora B和TRF1对白血病细胞增殖的影响。方法:以人髓原细胞白血病细胞株HL-60为研究对象,通过调节胞内CDK1表达来研究TRF1表达及其变化对细胞增殖和周期的影响。实验分为对照、shRNA-NC、CDK1-shRNA、pcDNA和pcDNA-CDK1共5组。采用RT-PCR检测各组细胞中CDK1表达;集落形成检测细胞增殖;Western blot检测CDK1、PLK1、Aurora B、TRF1及周期蛋白p53、p27和cyclinA的表达。结果:与对照组相比,CDK1-shRNA组中PLK1、Aurora B磷酸化水平及TRF1表达显著下调(P<0.05),细胞集落形成率显著下降,周期相关蛋白p53和p27表达显著升高而cyclinA表达则显著下降(P<0.05)。这表明,干扰CDK1表达可抑制HL-60细胞增殖并延长其进入有丝分裂的时间,从而延长细胞周期。而与对照组相比,pcDNA-CDK1组中CDK1过表达则使PLK1、Aurora B磷酸化水平及TRF1表达显著上升(P<0.05),克隆形成率显著升高,周期相关蛋白p53和p27显著下降且cyclinA表达显著上调(P<0.05)。这表明,CDK1过表达可刺激不良反应发生,从而促进HL-60细胞的增殖并缩短细胞周期。结论:敲除CDK1可抑制PLK1和Aurora B磷酸化并负向调节TRF1,从而抑制白血病细胞的增殖。Objective: To investigate the effect of CDK1 interference regulation of PLK1, Aurora B and TRF1 on the proliferation of leukemia cells. Methods: The human myelogenous leukemia cell line HL-60 was selected as the research object, and the effect of TRF1 expression and its changes on cell proliferation and cycle was investigated by regulating intracellular CDK1 expression. The objects were divided into 5 groups, including control group, shRNA-NC group,CDK1-shRNA group, pcDNA group and pcDNA-CDK1 group. RT-PCR was used to detect the CDK1 expression of cells in each group;colony formation was used to detect the proliferation of the cells. Western blot was used to detect the expression of CDK1, PLK1, Aurora B, TRF1, and cyclin p53, p27, cyclinA. Results: The phosphorylation level of PLK1,Aurora B and the expression of TRF1 in the CDK1-shRNA group were significantly down-regulated as compared with those in the control group(P<0.05). Compared with the control group, the cells in CDK1-shRNA group showed lower clone formation rate, the increasing of cycle-associated proteins p53 and p27 and the decreasing of cyclinA expression(P<0.05). It was shown that interfered CDK1 expression could inhibit the proliferation of HL-60 cells and prolong the time that they enter mitosis, thereby extending the cell cycle. Compared with the control group, the overexpressed CDK1 in the pcDNA-CDK1 group made the phosphorylation level of PLK1, Aurora B, and TRF1 expression increase significantly(P<0.05), also the colony formation rate(P<0.05). The cycle-related proteins p53 and p27 was downregulated,while cyclinA expression was up-regulate significantly(P<0.05). The results indicted that overexpressed CDK1 could stimulate adverse reactions, thereby promoting the proliferation of HL-60 cells and shortening the cell cycle.Conclusion: Knocking out CDK1 can inhibit the phosphorylation of PLK1 and Aurora B and negatively regulate TRF1,thereby inhibiting the proliferation of leukemia cells.

关 键 词：CDK1 有丝分裂 端粒酶 细胞周期 白血病 

分 类 号：R733.7[医药卫生—肿瘤]