分子生物学
生物
转铁蛋白受体
红细胞生成
K562细胞
流式细胞术
基因敲除
血红素
细胞分化
细胞培养
细胞
血红素
内科学
生物化学
基因
贫血
医学
遗传学
酶
作者
Zhan Gao,Jun Xiao,Xiaowei Li,Cuiying Li
出处
期刊:PubMed
日期:2022-06-01
卷期号:38 (6): 494-500
摘要
Objective To investigate the role of endothelial PAS domain-containing protein 1 (EPAS1) gene encoded hypoxia-inducible factor 2α (HIF-2α) in erythroid differentiation of K562 human erythroleukemia cells under hypoxic conditions. Methods K562 cells were treated with 40 μmol/L of hemin and 0.1 ng/mL of cytarabine for erythroid differentiation. After normoxic and hypoxic(50 mL/L O2) incubation, the ratio of CD235a+CD71+ cells was detected by flow cytometry. The percentage of hemoglobin-positive cells was detected by benzidine staining. The level of cell proliferation was detected by CCK-8 assay. The mRNA and protein levels of EPAS1, insulin receptor substrate 2 (IRS2) and γ-globin were detected by real-time quantitative PCR and Wester blot analysis. Besides, the changes in the erythroid differentiation of K562 cells were evaluated after knockdown of EPAS1. Results Hypoxia promoted the erythroid differentiation of K562 cells and upregulated the expression of EPAS1. After EPAS1 kncokdown, the ratio of CD235a+CD71+ cells and hemoglobin-positive cells decreased, and the expressions of IRS2 and γ-globin declined significantly. Conclusion Hypoxia can significantly up-regulate the expression of EPAS1 in K562 cells and promote the erythroid differentiation.
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