miR-34a-5p抑制K562细胞红系分化

miR-34a-5p inhibits the erythroid differentiation of K562 cells

目的探索miR-34a-5p对K562细胞红系分化的影响。方法分别用miR-34a-5p模拟物和反义抑制寡核苷酸转染K562细胞,用real-time PCR法检测过表达或干扰效率,并进一步用流式细胞术和联苯胺染色法检测K562细胞向红系的分化情况;通过Western blot方法检测miR-34a-5p的靶基因。结果 miR-34a-5p在K562细胞红系分化过程中呈现表达下降趋势;在K562细胞中过表达miR-34a-5p可抑制hemin诱导的红系分化(P<0.05);反之,干扰K562内源的miR-34a-5p表达会对K562红系分化产生促进作用(P<0.01);另一方面,miR-34a-5p通过靶向抑制c-MYB的表达抑制细胞向红系分化。结论 miR-34a-5p通过抑制c-MYB在K562细胞早期红系分化过程中发挥促进作用。

Objective To study the effects of microRNA-34a-5p on erythroid differentiation of K562 cells. Methods K562 cells were transfected with the microRNA-34a-5p mimics and antisense inhibitors specifically targeting microRNA-34a-5p,respectively. The effects of over-expression or knocking-down of microRNA-34a-5p were examined by Quantitative RT-PCR. Flow cytometry was performed to detect specific surface marker of erythroid cells.The benzidine staining assay was used to access the differentiation of K562 cells. Western blot was performed to detect miRNA targets. Results microRNA-34a-5p was down-regulated at the early stage of K562 erythroid differentiation. Over-expression of microRNA-34a-5p in K562 cells attenuates erythroid differentiation,in contrast,inhibition of microRNA-34a-5p accelerates erythroid pheotypes in K562 cells. c-MYB was found to be the direct target of microRNA-34a-5p in erythroid cells. Conclusions microRNA-34a-5p regulates early erythroid differentiation of K562 cells via repressing c-MYB.