Effect of MiR-451a on Erythroid Differentiation of K562 Cells under Hypoxia.
10.19746/j.cnki.issn.1009-2137.2020.06.045
- Author:
Cai-Yan HU
1
;
Hui-Jie ZHANG
1
;
Cheng-Bing FU
1
;
Fang LIU
2
Author Information
1. Department of Biochemistry, Medical College of Qinghai University, Xining 810000, Qinghai Province, China.
2. Department of Biochemistry, Medical College of Qinghai University, Xining 810000, Qinghai Province, China,E-mail: qhxnlf2006@163.com.
- Publication Type:Journal Article
- MeSH:
14-3-3 Proteins;
Cell Differentiation;
Erythroid Cells/metabolism*;
GATA1 Transcription Factor/metabolism*;
Humans;
Hypoxia;
K562 Cells;
MicroRNAs/genetics*
- From:
Journal of Experimental Hematology
2020;28(6):2071-2078
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the changes of GATA-1 protein expression during erythroid differentiation of K562 cells under hypoxia and how GATA-1 can regulate erythroid differentiation by up-regulating the expression of miR-451a and inhibiting the expression of 14-3-3ζ.
METHODS:K562 cells were divided into 2 groups: the normoxia group and the hypoxia group, after the induction of hemin for 96 h, the positive cells rate of the benzidine staining, the mRNA expression of γ-globin and the expression of CD235a were detected, and the success of the model was verified. The changes of GATA-1 and miR-451a expression in the above-mentioned 2 groups, the changes of miR-451a expression after over-expressed GATA-1 were detected by Western blot and qRT-PCR. The cells in normoxic group and hypoxia group were divided into negative control group (NC group) and miR-451a over-expression group respectively, and the degree of erythroid differentiation in the four groups was judged according to the corresponding erythroid differentiation indexes, and the expression of 14-3-3ζ was detected by Western blot after over-expressed miR-451a.
RESULTS:The positive cell rate of benzidine staining, mRNA expression of γ-globin and the expression of CD235a after 96 h induction by K562 cells under hypoxia were significantly higher than 0 h, suggesting that the erythroid differentiation model of K562 cells under hypoxia was replicated successfully. The expression levels of GATA-1 protein and miR-451a in the hypoxic group were significantly higher than that in the normoxic group (P<0.05). The expression level of miR-451a in hypoxia group was significantly higher than that in NC group after overexpressed GATA-1 (P<0.05). After over-expressed of miR-451a under hypoxia, the positive cell rate of benzidine staining, the mRNA expression level of γ-globin and the expression of CD235a were significantly higher than those in NC group (P<0.05). The expression level of 14-3-3ζ protein in miR-451a over-expressed group was lower than that in NC group under hypoxia (P<0.05).
CONCLUSION:Hypoxia can significantly increase the expression of GATA-1 protein, and the increase of GATA-1 expression can up-regulate the expression of miR-451a, thereby inhibiting the expression of 14-3-3ζ protein, which hinders the cell proliferation in erythroid differentiation model of K562 cells and plays an important role in promoting erythroid differentiation.
