摘要
目的:研究RNA结合蛋白ABCF1在人急性髓系白血病(acute myeloid leukemia,AML)发生发展过程中的功能。方法:以临床初诊的AML患者为实验组,正常人为对照组,采用RT-qPCR检测ABCF1 mRNA在正常人外周血中单个核细胞(peripheral blood mononuclear cells,PBMCs)和AML患者骨髓有核细胞中的表达;用针对ABCF1的2个shRNA慢病毒颗粒感染人单核细胞白血病细胞株THP-1,用CCK-8法检测THP-1细胞的增殖,用PI、Annexin V染色结合流式细胞仪检测细胞凋亡和细胞周期。利用ss-m^(6)A和ss-A探针进行RNA pull-down实验,分析ABCF1与m^(6)A的结合能力。结果:ABCF1在实验组AML患者中的表达明显高于对照组(P<0.01),在THP-1细胞中抑制内源ABCF1的表达能够促进细胞凋亡(P<0.05),抑制细胞增殖(P<0.001),同时明显减少S期细胞的比例。与ss-A探针相比,ABCF1蛋白具有特异的m^(6)A结合能力。结论:ABCF1在AML细胞来源的THP-1细胞中发挥原癌基因作用,调控THP-1细胞的生理功能。
Objective:To explore the function of RNA-binding protein ABCF1 in the development of human acute myeloid leukemia(AML).Methods:The clinically diagnosed AML patients were enrolled in the experimental group,and normal people in the control group.The expression level of ABCF1 mRNA was respectively detected by RT-qPCR in peripheral blood mononuclear cells(PBMCs)of normal people and bone marrow nucleated cells of AML patients.Human monocytic leukemia cell line THP-1 was infected with two shRNA lentivirus particles targeting ABCF1.The proliferation of THP-1 cells was detected by CCK-8 analysis,and apoptosis and cell cycle were detected by PI and Annexin V staining combined with flow cytometry.RNA pull-down test was carried out by using ss-m^(6)A and ss-A probes,and the binding ability of ABCF1 and m^(6)A was analyzed.Results:The expression of ABCF1 in AML patients in experimental group was significantly higher than that in control group(P<0.01).ABCF1 knock-down significantly promoted apoptosis(P<0.05),and suppressed cell proliferation(P<0.001),as well as affected the cell cycle of THP-1 cells.Conclusion:ABCF1 plays an important role of proto-oncogene in THP-1 cells derived from AML cells and regulates the physiological functions of THP-1 cells.
作者
吴爽
罗文依
杨健
余佳
王东生
Wu Shuang;Luo Wenyi;Yang Jian;Yu Jia;Wang Dongsheng(School of Basic Medical Sciences,North Sichuan Medical College;Department of Laboratory Medicine,North Sichuan Medical College;Instituteof Basic Medical Sciences,Chinese Academy of Medical Sciences;Department of Laboratory Medicine,Sichuan Cancer Hospital&Institute)
出处
《重庆医科大学学报》
CAS
CSCD
北大核心
2022年第9期1025-1029,共5页
Journal of Chongqing Medical University
基金
四川省科技厅应用基础资助项目(编号:2016JY0171)。
