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miR‑let‑7c‑3p targeting on Egr‑1 contributes to the committed differentiation of leukemia cells into monocyte/macrophages

  • Authors:
    • Fu Qi
    • Xinping Wang
    • Shouzhen Zhao
    • Chaozhe Wang
    • Ruijing Sun
    • Huan Wang
    • Pengchao Du
    • Jing Wang
    • Xidi Wang
    • Guosheng Jiang

  • View Affiliations

  • Published online on: June 22, 2022     https://doi.org/10.3892/ol.2022.13393
  • Article Number: 273

  • Copyright: © Qi et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

In preliminary experiments, it was found that the expression of early growth response‑1 (Egr‑1) was upregulated during the committed differentiation of leukemia cells into monocytes/macrophages. The cross‑analysis of gene chip detection and database prediction indicated that Egr‑1 was associated with upstream microRNA (miR)‑let‑7c‑3p, thus the present study focused on the role of the miR‑let‑7c‑3p/Egr‑1 signaling axis in the committed differentiation of leukemia cells into monocytes/macrophages. Phorbol 12‑myristate 13‑acetate (PMA) was used to induce the directed differentiation of human K562 leukemia cells into monocytes/macrophages and the differentiation of K562 leukemia cells was determined by cell morphology observation and expression of differentiation antigens CD11b and CD14 by flow cytometry. The expression levels of Egr‑1 and miR‑let‑7c‑3p were detected by reverse transcription‑quantitative PCR and the protein expression of Egr‑1 was detected by western blotting. The effect of Egr‑1 on the differentiation of K562 cells was detected by short interfering (si)RNA interference assay. A dual‑luciferase reporter assay was used to detect target binding of miR‑let‑7c‑3p on the 3'UTR of Egr‑1. Cell transfection of miR‑let‑7c‑3p mimics and inhibitors was used to modulate the expression of miR‑let‑7c‑3p, as indicated by RT‑qPCR assays. Western blotting was also used to examine the effect of miR‑let‑7c‑3p on Egr‑1 expression. The PMA‑induced differentiation of K562 cells was transfected with miR‑let‑7c‑3p and the expression of differentiation antigen was detected by flow cytometry. A differentiation model of K562 leukemia cells into monocytes/macrophages was induced by PMA, which was indicated by morphological observations and upregulation of CD11b and CD14 antigens. The gene or protein expression of Egr‑1 was significantly higher compared with that of the control group, while the expression of miR‑let‑7c‑3p was significantly lower compared with that of the control group. siRNA interference experiments showed that the expression of cell differentiation antigen CD14 in the 100 µg/ml PMA + si‑Egr‑1 group was significantly lower compared with that in the 100 µg/ml PMA + si‑ctrl group. The dual luciferase reporter gene results showed that the luciferase activity of the co‑transfected mimic and Egr‑1 WT groups was significantly lower than that of the NC control group, while the luciferase activity of the co‑transfected mimic and Egr‑1 MUT groups was comparable to that of the NC control group. Therefore, the dual‑luciferase reporter gene assay confirmed that miR‑let‑7c‑3p can target Egr‑1. Western blotting showed that the expression of Egr‑1 following transfection with miR‑let‑7c‑3p inhibitor was significantly higher compared with that of the negative control and the expression of Egr‑1 after transfection with miR‑let‑7c‑3p mimic was significantly lower than that of the negative control. Following exposure to PMA, the expressions of CD11b and CD14 in the miR‑let‑7c‑3p inhibitor group were significantly higher than those in the miR‑let‑7c‑3p NC group, as indicated by CD11b and CD14 respectively. In conclusion, miR‑let‑7c‑3p could bind to the 3'UTR of Egr‑1 and negatively regulated Egr‑1 expression. The miR‑let‑7c‑3p/Egr‑1 signaling axis was closely associated with the committed differentiation of K562 cells from leukemia cells to monocytes/macrophages.
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August-2022
Volume 24 Issue 2

Print ISSN: 1792-1074
Online ISSN:1792-1082

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Spandidos Publications style
Qi F, Wang X, Zhao S, Wang C, Sun R, Wang H, Du P, Wang J, Wang X, Jiang G, Jiang G, et al: miR‑let‑7c‑3p targeting on Egr‑1 contributes to the committed differentiation of leukemia cells into monocyte/macrophages. Oncol Lett 24: 273, 2022.
APA
Qi, F., Wang, X., Zhao, S., Wang, C., Sun, R., Wang, H. ... Jiang, G. (2022). miR‑let‑7c‑3p targeting on Egr‑1 contributes to the committed differentiation of leukemia cells into monocyte/macrophages. Oncology Letters, 24, 273. https://doi.org/10.3892/ol.2022.13393
MLA
Qi, F., Wang, X., Zhao, S., Wang, C., Sun, R., Wang, H., Du, P., Wang, J., Wang, X., Jiang, G."miR‑let‑7c‑3p targeting on Egr‑1 contributes to the committed differentiation of leukemia cells into monocyte/macrophages". Oncology Letters 24.2 (2022): 273.
Chicago
Qi, F., Wang, X., Zhao, S., Wang, C., Sun, R., Wang, H., Du, P., Wang, J., Wang, X., Jiang, G."miR‑let‑7c‑3p targeting on Egr‑1 contributes to the committed differentiation of leukemia cells into monocyte/macrophages". Oncology Letters 24, no. 2 (2022): 273. https://doi.org/10.3892/ol.2022.13393