MiR-152-3p promotes the development of chronic myeloid leukemia by inhibiting p27

L. Wang, Y. Wang, J. Lin

Department of Hematology, the Affiliated Yantai Yuhuangding Hospital of Qingdao University, Yantai, China. junlin330@sohu.com

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• Hematology

OBJECTIVE: The aim of this study was to investigate the relationship between micro ribonucleic acid (miR)-152-3p and chronic myeloid leukemia (CML), and to explore the underlying mechanism.

PATIENTS AND METHODS: The expression level of miR-152-3p in the bone marrow of 40 CML patients and 30 normal controls was detected by fluorescence quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). MiR-152-3p was up-regulated or down-regulated in the CML cell line (K562) by transfection with overexpressed lentivirus LV5-miR-152-3p or interfered lentivirus LV3-miR-152-3p, respectively. Transfection efficiency of miR-152-3p was detected by qRT-PCR. Cell counting kit-8 (CCK-8) assay was applied to measure the proliferation ability of K562 cells. Flow cytometry was used to assess the cell cycle and apoptosis rate of K562 cells after transfection. P27Kip1 (p27) was confirmed as the potential target of miR-152-3p by on-line target gene prediction software. Moreover, the interaction between miR-152-3p and p27 was analyzed by luciferase reporter gene assay and Western blot.

RESULTS: MiR-152-3p was highly expressed in the bone marrow of CML patients and cell lines. In vitro experiments indicated that the apoptosis rate of K562 cells in the lentivirus LV3-miR-152-3p interference group was significantly increased, and the cell cycle was arrested in G0G1 phase. Meanwhile, the proliferation of K562 cells was markedly inhibited. However, LV5-miR-152-3p transfection remarkably promoted the proliferation and cell cycle progression of K562 cells. We searched three online public databases to predict the potential target of miR-152-3p and found that p27 was a direct target of miR-152-3p. Luciferase reporter gene assay and Western blot confirmed our hypothesis. In addition, subsequent experiments showed that up-regulation of p27 attenuated the effect of miR-152-3p on the ability of CML cells.

CONCLUSIONS: MiR-152-3p was highly expressed in the bone marrow of CML patients and cells. Meanwhile, miR-152-3p inhibited the expression of its downstream protein p27, thus promoting the proliferation of K562 cells and the entry of cells into the cell cycle. In addition, it inhibited cell apoptosis, which might be a potential target for the incidence and development of CML.

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