Di-san junyi daxue xuebao (Nov 2021)

MiRNA-495 regulates proliferation and apoptosis of acute myeloid leukemia cells with MLL gene rearrangement by targeting STYX

  • YANG Rui,
  • NING Jing,
  • JING Lijuan,
  • WANG Hainan,
  • CUI Lijuan

DOI
https://doi.org/10.16016/j.1000-5404.202105091
Journal volume & issue
Vol. 43, no. 22
pp. 2407 – 2413

Abstract

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Objective To investigate the effect of miRNA-495 on the proliferation and apoptosis of acute myeloid leukemia (AML) cells with mixed lineage leukemia (MLL) gene rearrangement. Methods Ten patients diagnosed with AML with MLL-AF9 fusion gene positive (6 males, 4 females, mean age of 39.37±2.71 years), and 10 cases of AML (5 males, 5 females, mean age of 41.12±1.92 years) admitted in our hospital from August 2017 to May 2019 were enrolled in this study, and another 10 patients (6 males, 4 females, mean age of 42.31±3.21 years) who attended the surgical outpatient department during the same period were also collected as the control. THP-1 cells (containing MLL-AF9 fusion gene) were divided into miRNA-495 and control groups based on transfection with miRNA-495 or not, and the expression of miRNA-495 in each group was detected by RT-qPCR; CCK-8 assay was used to determine the proliferation of transfected THP-1 cells in each group; RT-qPCR and Western blotting were performed to measure the expression of proliferation-related factors (Ki-67 and PCNA) and apoptosis-related factors (Bcl-2, Bax and Caspase-3). The target genes of miRNA-495 were predicted by TargetScan and then verified by dual-luciferase reporter assay. Finally, RT-qPCR and Western blotting were adopted to detect the expression of serine/threonine/tyrosine interacting protein (STYX) in groups as follows: miRNA-495 group, control group, STYX group (transfected with STYX) and miRNA-495+STYX group (transfected with miRNA-495 and STYX). Results MiRNA-495 expression was down-regulated in AML patients with MLL-AF9 fusion gene, with relative expression of 0.25±0.08 (95%CI=-0.86~-0.65, P < 0.001); miRNA-495 inhibited cell proliferation (P < 0.001) and induced apoptosis of THP-1 cells, and it targetedly bound to STYX (P < 0.001). In addition, STYX overexpression reversed the effects of miRNA-495 on THP-1 cell proliferation and apoptosis. Conclusion MiRNA-495 regulates the proliferation and apoptosis of AML cells with MLL gene rearrangement by targeting STYX.

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