MiR-10b regulates the proliferation and apoptosis of pediatric acute myeloid leukemia through targeting HOXD10

C.-J. Wang, H. Zou, G.-F. Feng

Laboratory Medicine, Wendeng Orthopedics Hospital of Shandong Province, Wendeng, China. shixianhuiqing@163.com

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• Hematology

OBJECTIVE: To investigate the role of miR-10b in the proliferation and apoptosis of acute myeloid leukemia (AML), and to explore the underlying mechanism.

PATIENTS AND METHODS: The expression level of miR-10b in clinical AML cases and cell lines was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The interaction between miR-10b and homeobox D10 (HOXD10) was confirmed by qRT-PCR, Western blotting and Luciferase assay. The effect of miR-10b on biological functions of AML cell line (HL60) was analyzed in vitro. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay and colony formation assay were used to detect the proliferation and colony formation ability of AML cells, respectively. Meanwhile, flow cytometry and TUNEL staining were applied to measure cell cycle and apoptosis of AML cells, respectively.

RESULTS: miR-10b was significantly up-regulated in AML cases and cell lines. The potential target genes of miR-10b were analyzed by three public databases. Results showed that HOXD10 was a direct target of miR-10b. QRT-PCR, Western blotting and luciferase assay confirmed the regulatory effect of miR-10b on HOXD10. Overexpression of miR-10b accelerated the proliferation and colony formation ability of AML cells. Meanwhile, miR-10b overexpression decreased the percentage of AML cells in the G0/G1 phase when compared with S phase, and suppressed the apoptosis of AML cells. However, the addition of HOXD10 could reverse the effects of miR-10b.

CONCLUSIONS: MiR-10b could regulate the proliferation, colony formation, cell cycle and apoptosis of AML cells through targeting HOXD10, indicating that miR-10b might be a potential therapeutic target for the treatment of AML.

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