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。
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.
出处
《生物工程学报》
CAS
CSCD
北大核心
2013年第7期946-954,共9页
Chinese Journal of Biotechnology
基金
国家自然科学基金(No.31201103)资助~~
