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Enforced miR-144-3p Expression as a Non-Invasive Biomarker for the Acute Myeloid Leukemia Patients Mainly by Targeting NRF2 by Xuelin Sun, Dejun Liu, Yuan Xue, Xin Hu

Background: MicroRNAs (miRNAs) are endogenous non-coding RNAs of 19-25 nucleotides in length. Abnormal miRNA expression has been identified in various types of cancer. However, the aberrant expression of miR-1244 has not been reported in acute myeloid leukemia.
Methods: First, the expression of miR-144-3p was explored in the bone marrow and peripheral blood of AML patients and healthy control. Then, we also evaluated the level of miR-144-3p in HL-60 cells. The possible target gene of miR-144-3p was predicted using TargetScan. Dual luciferase reporter assay was applied to validate the target gene of miR-144-3p. Cell viability and apoptosis on miR-144-3p was explored. Western blot analysis was applied to identify the downstream signaling of miR-144-3p.
Results: Our data showed that miR-144-3p was markedly increased in both the peripheral blood and bone marrow of AML patients compared with healthy controls. Moreover, we also found an increased expression of miR144-3p in HL-60 cells. In comparison, NRF2 protein expression was significantly decreased in HL-60 cells. Dual luciferase reporter assays demonstrated that miR-144-3p significantly suppressed the relative luciferase reporter activity of a pmirGLO-NRF2-3’UTR. In accordance with the downstream effects of NRF2 overexpression, inhibition of miR-144-3p reduced cell viability and prompted apoptosis. More importantly, we found that the inhibition of miR-144-3p in HL-60 cells could not enhance Caspase-3 activation when NRF2 protein expression was silenced.
Conclusions: These findings suggest a potential oncogenic function of miR-144-3p in HL-60 cells, which is mainly achieved by targeting NRF2.

DOI: 10.7754/Clin.Lab.2016.161116