Abstract
Purpose
Abnormal expression of miRNAs is intimately related to a variety of human cancers. The purpose of this study is to confirm the expression of miR-181a and elucidate its physiological function and mechanism in pediatric acute myeloid leukemia (AML).
Methods
Pediatric AML patients and healthy controls were enrolled, and the expression of miR-181a and ataxia telangiectasia mutated (ATM) in tissues were examined using quantitative PCR. Moreover, cell proliferation and cell cycle were evaluated in several cell lines (HL60, NB4 and K562) by using flow cytometry after transfected with miR-181a mimics and inhibitors, or ATM siRNA and control siRNA. Finally, ATM as the potential target protein of miR-181a was examined.
Results
We found that miR-181a was significantly increased in pediatric AML, which showed an inverse association with ATM expression. Overexpressed miR-181a in cell lines significantly enhanced cell proliferation, as well as increased the ratio of S-phase cells by miR-181a mimics transfection in vitro. Luciferase activity of the reporter construct identified ATM as the direct molecular target of miR-181a. ATM siRNA transfection significantly enhanced cell proliferation and increased the ratio of S-phase cells in vitro.
Conclusion
The results revealed novel mechanism through which miR-181a regulates G1/S transition and cell proliferation in pediatric AML by regulating the tumor suppressor ATM, providing insights into the molecular mechanism in pediatric AML.
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Acknowledgments
This work was supported by National Nature and Science Grant of China (81272310).
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The authors declare that they have no conflict of interest.
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Xiaodan Liu, Wang Liao and Hongxia Peng authors have contributed equally to the study.
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Supplement Fig. 1
The relationship of miR-181a expression and prognosis in 27 AML patients with complete follow-up data. a miR-181a expression in patients who achieved a complete remission versus patients who developed distant relapse or death. b Relapse-free survival according to miR-181a expression levels in pediatric AML patients with high (above the median miR-181a expression value) or low (at or below the median miR-181a expression value) expression groups. Statistical differences between curves were calculated using the log-rank test (TIFF 10822 kb)
Supplement Table 1
Characteristics of pediatric AML patients (N = 57) (DOCX 17 kb)
Supplement Table 2
Primers and other oligonucleotides sequences (DOCX 14 kb)
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Liu, X., Liao, W., Peng, H. et al. miR-181a promotes G1/S transition and cell proliferation in pediatric acute myeloid leukemia by targeting ATM. J Cancer Res Clin Oncol 142, 77–87 (2016). https://doi.org/10.1007/s00432-015-1995-1
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DOI: https://doi.org/10.1007/s00432-015-1995-1