Carcinogenesis, Teratogenesis & Mutagenesis ›› 2024, Vol. 36 ›› Issue (6): 438-443.doi: 10.3969/j.issn.1004-616x.2024.06.003

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Regulatory role of the miR-520c-3p/ CXCL8 axis in erythroid differentiation of chronic myeloid leukemia cells

YANG Yu1, SHANG Yu1, NI Lei2, GONG Zilin1, WANG Ran3, WANG Dan4, WANG Jingyan2   

  1. 1. Key Laboratory of Microecology-Immune Regulatory Network and Related Diseases, Basic Medical College of Jiamusi University, Jiamusi 154007;
    2. The First Affiliated Hospital of Jiamusi University, Jiamusi 154003;
    3. Jiamusi Central Hospital, Jiamusi 154002;
    4. Clinical Medical College of Jiamusi University, Jiamusi 154003, Heilongjiang, China
  • Received:2024-08-21 Revised:2024-10-21 Published:2024-12-04

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Abstract: OBJECTIVE： To explore the role and mechanism of the miR-520c-3p/CXCL8 axis in erythroid differentiation of chronic myeloid leukemia (CML). METHODS： K562 cells were induced to differentiate with hemin and treated with CXCL8 (0，20，40，60，80 and 100 ng/mL) for 72 h. Expression of γ-globin mRNA was examined by RT-qPCR to analyze the effect of CXCL8 on erythroid differentiation. miR-520c-3p mimics and inhibitor were transfected into K562 cells to enhance or inhibit miR-520c-3p function，respectively. Expression of γ-globin mRNA was measured by RT-qPCR to assess the impact of miR-520c-3p on erythroid differentiation of K562 cells. Western blot and RT-qPCR experiments were performed to detect CXCL8 expression and observe the influence of miR-520c-3p on CXCL8 expression. The cells were divided into the experiment group (co-transfected with miR-520c-3p mimics and CXCL8 3'-UTR) and the control group (co-transfected with mimics-control and CXCL8 3'-UTR)，a dual luciferase reporter gene assay was conducted to examine the targeting relationship between miR-520c-3p and the 3'-UTR of CXCL8. RESULTS： Compared to the control group，expression of CXCL8 mRNA was significantly increased after hemin-induced differentiation (P<0.01). Compared to the untreated group，the γ-globin mRNA expression of CXCL8 treatment groups were significantly elevated (P<0.01). Compared with the negative control group，the expression levels of γ-globin mRNA，CXCL8 mRNA，and CXCL8 protein in K562 cells transfected with miR-520c-3p mimics were all decreased (P<0.01). In cells transfected with the miR-520c-3p inhibitor，expression levels of γ-globin mRNA，CXCL8 mRNA，and CXCL8 protein were all increased (P<0.01). The dual luciferase reporter gene assay revealed a significant decrease in relative fluorescence activities in the experimental group compared to the control group (P<0.01). CONCLUSION： This study reveals the key role of the miR-520c-3p/CXCL8 signaling axis in the erythroid differentiation of CML cells. miR-520c-3p exerted an inhibitory effect on the erythroid differentiation of K562 cells by targeting the expression of CXCL8.

Key words: chronic myeloid leukemia, miR-520c-3p, CXCL8, erythroid differentiation

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