RNA结合蛋白ABCF1在AML发生发展过程中的功能研究

Function of RNA-binding protein ABCF1 in the development of human acute myeloid leukemia

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-m6A和ss-A探针进行RNA pull-down实验，分析ABCF1与m6A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<0.0001），同时明显减少细胞周期S期细胞的比例。与ss-A探针相比，ABCF1蛋白具有特异的m6A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。

Abstract:

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-m6A and ss-A probes, and the binding ability of ABCF1 and m6A 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.