MicroRNAs 221 and 222 inhibit normal erythropoiesis and erythroleukemic cell growth via kit receptor down-modulation

Nadia Felli1, Laura Fontana1, Elvira Pelosi1, Rosanna Botta1, Désirée Bonci1, Francesco Facchiano1, Francesca Liuzzi1, Valentina Lulli1, Ornella Morsilli1, Simona Santoro1, M Valtieri1, George A. Calin1, Chang-Gong Liu1, Antonio Sorrentino1, Carlo M. Croce1, C Peschle1
1Department of Hematology, Oncology, and Molecular Medicine, Istituto Superiore di Sanità, 00161 Rome, Italy; Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA 19107; and Institute of Genetics Comprehensive Cancer Center, Ohio State University, Columbus, OH 43210

Tóm tắt

MicroRNAs (miRs) are small noncoding RNAs that regulate gene expression primarily through translational repression. In erythropoietic (E) culture of cord blood CD34+ progenitor cells, the level of miR 221 and 222 is gradually and sharply down-modulated. Hypothetically, this decline could promote erythropoiesis by unblocking expression of key functional proteins. Indeed, ( i ) bioinformatic analysis suggested that miR 221 and 222 target the 3′ UTR of kit mRNA; ( ii ) the luciferase assay confirmed that both miRs directly interact with the kit mRNA target site; and ( iii ) in E culture undergoing exponential cell growth, miR down-modulation is inversely related to increasing kit protein expression, whereas the kit mRNA level is relatively stable. Functional studies show that treatment of CD34+ progenitors with miR 221 and 222, via oligonucleotide transfection or lentiviral vector infection, causes impaired proliferation and accelerated differentiation of E cells, coupled with down-modulation of kit protein: this phenomenon, observed in E culture releasing endogenous kit ligand, is magnified in E culture supplemented with kit ligand. Furthermore, transplantation experiments in NOD-SCID mice reveal that miR 221 and 222 treatment of CD34+ cells impairs their engraftment capacity and stem cell activity. Finally, miR 221 and 222 gene transfer impairs proliferation of the kit+ TF-1 erythroleukemic cell line. Altogether, our studies indicate that the decline of miR 221 and 222 during exponential E growth unblocks kit protein production at mRNA level, thus leading to expansion of early erythroblasts. Furthermore, the results on kit+ erythroleukemic cells suggest a potential role of these miRs in cancer therapy.

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