Sun Xuelin, Liu Dejun, Xue Yuan, Hu Xin
Clin Lab. 2017 Apr 1;63(4):679-687. doi: 10.7754/Clin.Lab.2016.161116.
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.
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.
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.
These findings suggest a potential oncogenic function of miR-144-3p in HL-60 cells, which is mainly achieved by targeting NRF2.
微小RNA（miRNA）是长度为19 - 25个核苷酸的内源性非编码RNA。在各种类型的癌症中已发现miRNA表达异常。然而，miR - 1244在急性髓系白血病中的异常表达尚未见报道。
首先，研究了miR - 144 - 3p在急性髓系白血病患者骨髓和外周血以及健康对照中的表达。然后，我们还评估了HL - 60细胞中miR - 144 - 3p的水平。使用TargetScan预测miR - 144 - 3p的可能靶基因。应用双荧光素酶报告基因检测法验证miR - 144 - 3p的靶基因。探讨了miR - 144 - 3p对细胞活力和凋亡的影响。采用蛋白质免疫印迹分析来鉴定miR - 144 - 3p的下游信号传导。
我们的数据显示，与健康对照相比，急性髓系白血病患者外周血和骨髓中的miR - 144 - 3p明显升高。此外，我们还发现HL - 60细胞中miR144 - 3p的表达增加。相比之下，HL - 60细胞中NRF2蛋白表达显著降低。双荧光素酶报告基因检测表明，miR - 144 - 3p显著抑制pmirGLO - NRF2 - 3'UTR的相对荧光素酶报告基因活性。与NRF2过表达的下游效应一致，抑制miR - 144 - 3p可降低细胞活力并促使细胞凋亡。更重要的是，我们发现当NRF2蛋白表达沉默时，在HL - 60细胞中抑制miR - 144 - 3p并不能增强Caspase - 3的激活。
这些发现表明miR - 144 - 3p在HL - 60细胞中具有潜在的致癌功能，这主要是通过靶向NRF2实现的。
