title: Increase of microRNA-210, decrease of raptor gene expression and alteration of mammalian target of rapamycin regulated proteins following mithramycin treatment of human erythroid cells.
abstract: Expression and regulation of microRNAs is an emerging issue in erythroid differentiation and globin gene expression in hemoglobin disorders. In the first part of this study microarray analysis was performed both in mithramycin-induced K562 cells and erythroid precursors from healthy subjects or β-thalassemia patients producing low or high levels of fetal hemoglobin. We demonstrated that: (a) microRNA-210 expression is higher in erythroid precursors from β-thalassemia patients with high production of fetal hemoglobin; (b) microRNA-210 increases as a consequence of mithramycin treatment of K562 cells and human erythroid progenitors both from healthy and β-thalassemia subjects; (c) this increase is associated with erythroid induction and elevated expression of γ-globin genes; (d) an anti-microRNA against microRNA-210 interferes with the mithramycin-induced changes of gene expression. In the second part of the study we have obtained convergent evidences suggesting raptor mRNA as a putative target of microRNA-210. Indeed, microRNA-210 binding sites of its 3'-UTR region were involved in expression and are targets of microRNA-210-mediated modulation in a luciferase reporter assays. Furthermore, (i) raptor mRNA and protein are down-regulated upon mithramycin-induction both in K562 cells and erythroid progenitors from healthy and β-thalassemia subjects. In addition, (ii) administration of anti-microRNA-210 to K562 cells decreased endogenous microRNA-210 and increased raptor mRNA and protein expression. Finally, (iii) treatment of K562 cells with premicroRNA-210 led to a decrease of raptor mRNA and protein. In conclusion, microRNA-210 and raptor are involved in mithramycin-mediated erythroid differentiation of K562 cells and participate to the fine-tuning and control of γ-globin gene expression in erythroid precursor cells.
authors: Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
authors: Nicoletta
authors: N
authors: Bianchi
authors: Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy; Laboratory for the Development of Pharmacological and Pharmacogenomic Therapy of Thalassaemia, Biotechnology Center, Ferrara University, Ferrara, Italy.
authors: Alessia
authors: A
authors: Finotti
authors: Department of Morphology, Surgery and Experimental Medicine, Ferrara University, Ferrara, Italy; Laboratory for Technologies of Advanced Therapies (LTTA), Ferrara University, Ferrara, Italy.
authors: Manuela
authors: M
authors: Ferracin
authors: Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
authors: Ilaria
authors: I
authors: Lampronti
authors: Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
authors: Cristina
authors: C
authors: Zuccato
authors: Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
authors: Giulia
authors: G
authors: Breveglieri
authors: Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
authors: Eleonora
authors: E
authors: Brognara
authors: Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
authors: Enrica
authors: E
authors: Fabbri
authors: Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy.
authors: Monica
authors: M
authors: Borgatti
authors: Department of Morphology, Surgery and Experimental Medicine, Ferrara University, Ferrara, Italy; Laboratory for Technologies of Advanced Therapies (LTTA), Ferrara University, Ferrara, Italy.
authors: Massimo
authors: M
authors: Negrini
authors: Department of Life Sciences and Biotechnology, Ferrara University, Ferrara, Italy; Laboratory for the Development of Pharmacological and Pharmacogenomic Therapy of Thalassaemia, Biotechnology Center, Ferrara University, Ferrara, Italy.
authors: Roberto
authors: R
authors: Gambari
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000096:biosynthesis* / Q000235:genetics; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D041905:Erythroid Cells / Q000378:metabolism*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D020014:K562 Cells; D008297:Male; D035683:MicroRNAs / Q000096:biosynthesis* / Q000235:genetics; D008926:Plicamycin / Q000494:pharmacology*; D000076223:Regulatory-Associated Protein of mTOR; D058570:TOR Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D017086:beta-Thalassemia / Q000235:genetics / Q000378:metabolism; D055545:gamma-Globins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C543800:MIRN210 microRNA, human; D035683:MicroRNAs; C463659:RPTOR protein, human; D000076223:Regulatory-Associated Protein of mTOR; D055545:gamma-Globins; C546842:MTOR protein, human; D058570:TOR Serine-Threonine Kinases; D008926:Plicamycin