miR-24通过靶基因Sp1调控β-类珠蛋白表达

miR-24 improves β-like globin gene expression through targeting Sp1

摘要:

为了研究miR-24对于珠蛋白表达的调控作用，并明确其作用机制。首先采用定量PCR的方法确定miR-24在红系分化过程中的表达变化情况，以及miR-24过表达后珠蛋白的表达变化情况。进而通过报告基因实验以及Western blotting的方法确定miR-24的靶基因。通过表型回复实验证明miR-24是否通过靶基因调控珠蛋白的表达。结果发现在hemin诱导的K562细胞以及EPO诱导的造血干/祖细胞向红系分化过程中，miR-24表达上调，在K562细胞中过表达miR-24可以促进红系分化过程中ε-和γ-珠蛋白的表达上调，进一步的研究表明miR-24是通过靶基因Sp1来行使对珠蛋白的调控作用的。以上结果表明miR-24通过负调节其靶基因Sp1促进红系分化过程中珠蛋白的表达上调

关键词:miR-24，珠蛋白，Sp1，红系分化，K562细胞

Abstract:

We studied the function and mechanism of miR-24 in regulating β-like globin gene expression. We first detected the expression of miR-24 during erythroid differentiation and also detected the globin gene expression in miR-24 overexpressing K562 cells through q-PCR. Dual-luciferase reporter assay and Western blotting were used to identify target genes of miR-24. “Rescue experiment” was further used to investigate the regulation of miR-24 on globin gene expression whether depending on targeting Sp1 or not. We found that miR-24 increased during hemin-induced K562 cells and EPO-induced HPCs (hematopoietic progenitor cells) erythroid differentiation. Overexpression of miR-24 in K562 cells promoted the ε- and γ-globin gene expression during hemin-induced erythroid differentiation through targeting the negative globin regulator Sp1. These results suggested that miR-24 can improve the expression of β-like globin gene through targeting Sp1

Key words:miR-24, globin, Sp1, erythroid differentiation, K562 cells