MiR-27a Promotes Hemin-Induced Erythroid Differentiation of K562 Cells by Targeting CDC25B

Abstract

Background/Aims: MicroRNAs (miRNAs) play a crucial role in erythropoiesis. MiR-23a∼27a∼24-2 clusters have been proven to take part in erythropoiesis via some proteins. CDC25B (cell division control Cdc2 phosphostase B) is also the target of mir-27a; whether it regulates erythropoiesis and its mechanism are unknown. Methods: To evaluate the potential role of miR-27a during erythroid differentiation, we performed miR-27a gain- and loss-of-function experiments on hemin-induced K562 cells. We detected miR-27a expression after hemin stimulation at different time points. At the same time, the γ-globin gene also was measured via real-time PCR. According to the results of the chips, we screened the target protein of miR-27a through a dual-luciferase reporter assay and identified it via Western blot analyses. To evaluate the function of CDC25B, benzidine staining and flow cytometry were employed to detect the cell differentiation and cell cycle. Results: We found that miR-27a promotes hemin-induced erythroid differentiation of human K562 cells by targeting cell division cycle 25 B (CDC25B). Overexpression of miR-27a promotes the differentiation of hemin-induced K562 cells, as demonstrated by γ-globin overexpression. The inhibition of miR-27a expression suppresses erythroid differentiation, thus leading to a reduction in the γ-globin gene. CDC25B was identified as a new target of miR-27a during erythroid differentiation. Overexpression of miR-27a led to decreased CDC25B expression after hemin treatment, and CDC25B was up-regulated when miR-27a expression was inhibited. Moreover, the inhibition of CDC25B affected erythroid differentiation, as assessed by γ-globin expression. Conclusion: This study is the first report of the interaction between miR-27a and CDC25B, and it improves the understanding of miRNA functions during erythroid differentiation.

MiR‐873 inhibition enhances gefitinib resistance in non‐small cell lung cancer cells by targeting glioma‐associated oncogene homolog 1

“…In the human genome, miRNAs regulate protein encoding genes at a rate of approximately 1:3, controlling cell apoptosis, proliferation, differentiation, metabolism, individual development and tumorigenesis, and drug resistance . Target gene expression is regulated by two mechanisms: (i) binding to the untranslated region (3'UTR) of the target messenger RNA (mRNA) 3' end, inhibiting its translation; and (ii) like small interfering RNA (siRNA), miRNA binds to the target and degrades target mRNA . Recent studies have found that miRNAs regulate drug resistance by mediating their targeting genes in various cancers .…”

“…[14][15][16] Target gene expression is regulated by two mechanisms: (i) binding to the untranslated region (3'UTR) of the target messenger RNA (mRNA) 3' end, inhibiting its translation; and (ii) like small interfering RNA (siRNA), miRNA binds to the target and degrades target mRNA. [17][18][19] Recent studies have found that miRNAs regulate drug resistance by mediating their targeting genes in various cancers. [20][21][22][23][24] In recent years, abnormal expression of miR-873 has been found in many kinds of tumors, such as breast and ovarian cancers, and glioma.…”

FAM122A Inhibits Erythroid Differentiation through GATA1

“…Also, erythroid cells at successive terminal stages of human erythropoiesis were also identified by using the combination of glycophorin A (GPA) and Band 3 ( Auffray et al., 2001 ; Hu et al., 2013 ). Toward this end, we used hemin at 50 μM to treat human erythroleukemic K562 cells as a cellular model for erythroid differentiation induction, as indicated by benzidine staining-positive (DAB + ) cells (an indicator for Hb production) and accumulation of HbG ( Rutherford et al., 1979 ; Wang et al., 2018 ). More intriguingly, our results demonstrated that FAM122A mRNA and protein levels were gradually decreased upon hemin-induced erythroid differentiation ( Figures S1 A, 1 B, and 1C).…”

MicroRNAs as Potential Markers Involved in Erythroid Differentiation: A Systematic Literature Review

“…Furthermore, KLF3 could suppress the expression of miR-23a27a24-2 cluster (named as miR-23a cluster) by binding to the CACCC sites in the promoter of miR-23a cluster, miR-23a cluster and KLF3 formed a positive feedback loop to promote β-like globin expression during erythropoiesis [40]. Meanwhile, miR-27a promotes hemin-induced erythroid differentiation of K562 cells by targeting CDC25B [41].…”