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