Up-regulation of miR-125a-5p inhibits proliferation and invasion of acute myeloid leukemia cell lines

Abstract

Objective To find the regulatory effect of miR-125a-5p on the growth and invasion of acute myeloid leukemia (AML) cell lines. Methods The expression of miR-125a-5p and TRIM71 in bone marrow specimens of AML patients, AML cell lines (U937 and HL60) and PBMC were examined by RT-qPCR. The U937 and HL60 cells were divided into control group (control), miR-125a-5p mimic group (miR-125a-5p mimic), miRNA negative control group (miR-NC), TRIM71 over-expression plasmid group (pcDNA3.1-TRIM71) and control plasmid group(pcDNA3.1-NC). U937 and HL60 cells were transfected with miR-125a-5p mimic, pcDNA3.1-TRIM71 and negative control using Lipofectamine 2000. Cell proliferation was measured by MTT assay; Cell invasion was measured by Matrigel invasion assay; And cell apoptosis was detected by flow cytometry. The expression of TRIM71, Bax, Bcl-2, NF-κB p65 was detected by RT-qPCR or Western blot. The targeted regulation relationship between miR-125a-5p and TRIM71 was verified by luciferase reporter gene assay. Results Compared with healthy controls or PBMC, miR-125a-5p expression levels in bone marrow samples of AML patients and AML cell lines were significantly reduced (P＜0.05). The proliferation and invasion of U937 and HL60 cells in miR-125a-5p mimic group were significantly reduced, while the apoptosis rate was significantly increased (P＜0.05). Compared with control group, the proliferation and invasion of U937 and HL60 cells in pcDNA3.1-TRIM71 group significantly were increased(P＜0.05). Luciferase reporter gene assay detection showed that miR-125a-5p mimic effectively inhibited the luciferase activity of pGL3-TRIM71-WT. Compared with miR-125a-5p mimic group, the cell viability and cell invasion rate of (miR-125a-5p mimic+pcDNA3.1-TRIM71) group were increased significantly, while the apoptosis rate was decreased significantly(P＜0.05). The expression level of nucleus NF-κB p65 protein in (miR-125a-5p mimic+pcDNA3.1-TRIM71) group was significantly increased(P＜0.05). Conclusions Up-regulation of miR-125a-5p may inhibit proliferation and invasion of AML cell lines and induce apoptosis by targeting at TRIM71. In addition, miR-125a-5p can inhibit the proliferation of AML cells by inhibiting the activation of the NF-κB signaling pathway.

Keywords

• acute myeloid leukemia|mir-125a-5p|proliferation|invasion|apoptosis|nf-κb signaling pathway