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MicroRNA-193a represses c-kit expression and functions as a methylation-silenced tumor suppressor in acute myeloid leukemia

Oncogene volume 30, pages 3416–3428 (2011)Cite this article

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Abstract

Aberrant activation of c-kit proto-oncogene contributes to abnormal cell proliferation by altering the tyrosine kinase signaling and constitutes a crucial impetus for leukemogenesis. Epigenetic silencing of tumor-suppressive microRNAs (miRNAs) is a key oncogenic mechanism for the activation of oncogenes in tumors. In this study, several miRNAs potentially binding to the 3′-untranslated region of human c-kit mRNA were screened by luciferase reporter assays. Among these miRNAs, miR-193a was embedded in a CpG island and epigenetically repressed by promoter hypermethylation in acute myeloid leukemia (AML) cell lines and primary AML blasts, but not in normal bone marrow cells. Importantly, miR-193a levels were inversely correlated with c-kit levels measured in 9 leukemia cell lines and 27 primary AML samples. Restoring miR-193a expression in AML cells harboring c-kit mutation and/or overexpression, either by synthetic miR-193a transfection or by DNA hypomethylating agent 5-azacytidine (5-aza) treatment, resulted in a significant reduction in c-kit expression at both RNA and protein levels and inhibition of cell growth. The growth-inhibitory activity of miR-193a was associated with apoptosis and granulocytic differentiation. Moreover, 5-aza-induced c-kit reduction could be partially blocked by miR-193a inhibitor, leading to a reversal of antiproliferative and proapoptotic effects of 5-aza. These data reveal a critical role for methylation-repressed miR-193a in myeloid leukemogenesis and the therapeutic promise of upregulating miR-193a expression for c-kit-positive AML.

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Acknowledgements

We thank Y Ding, Y-Y Xu, N Wang, C-W Xu, M Li, M-X Chen and W-D Han for excellent technical assistance. We also thank Professor X-F Zheng and H-J Fu from Beijing Institute of Radiation Medicine, China, for presenting the modified pGL3-control plasmid and miRNA expression plasmids. This work was supported by the National Natural Science Foundation of China (nos 81000221, 90919044 and 30971297) and the National 973 Project of China (no. 2005CB522400).

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  1. X-N Gao and J Lin: These authors contributed equally to this work.

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  1. Department of Hematology, Chinese PLA General Hospital, Beijing, China

    X-N Gao, Y-H Li, L Gao, X-R Wang, W Wang, H-Y Kang, L-L Wang & L Yu

  2. Basic Medical Institute, Chinese PLA General Hospital, Beijing, China

    J Lin & G-T Yan

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Gao, XN., Lin, J., Li, YH. et al. MicroRNA-193a represses c-kit expression and functions as a methylation-silenced tumor suppressor in acute myeloid leukemia. Oncogene 30, 3416–3428 (2011). https://doi.org/10.1038/onc.2011.62

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