Hsa-Mir-625-5p Upregulation Promotes Apoptosis in AML Cell Line by Targeting ILK Pathway

Background: Growing evidence has demonstrated that micro-RNAs (miRNAs) have a major effect on development of differ-ent types of cancer including AML (Acute Myeloid Leukemia). Expression of miR-625 has decreased in acute myeloid leuke-mia cell lines.Therefore it seemed that overexpression of miR-625 could decrease tumorigenesis of AML cell lines through ILK (Integrin Linked Kinase) signaling pathway and reducing the associated oncogenes. The aim of this study is to evaluate the effect of hsa-miR-625-5p upregulation on apoptosis and proliferation of KG-1 cell line via ILK signaling pathway.Materials and Methods: The KG-1 cell line was transfected with pLenti-III-pre mir625-GFP through viral method. Then, overexpression of miR-625-5p as well as the expression level of ILK, AKT, GSK3, c-fos, NF-κB, Caspase 3, Cyclin D1 and stemness genes including KLF-4, OCT-4 and Nanog were ana-lyzed by quantitative PCR (qPCR). Western blotting was used to evaluate of NF-κB, Caspase 3 and p-β-catenin at the pro-tein level. Apoptosis was investigated by Annexin V and flow cytometry. Cell cycle analysis with PI and CCK-8 assay were performed to evaluate proliferation.Results: Flow cytometric analysis results of KG-1 cells trans-fected with pLenti-III-pre mir625-GFP construct showed a sig-nificant increase in cell apoptosis but no significant alteration in cell cycle. The CCK-8 assay demonstrated that hsa-miR-625-5p doesn’t affect proliferation. Gene expression of ILK and NF-κB were downregulated and AKT, c-fos, Caspase3, Cyclin D1, KLF-4, OCT-4 and Nanog were upregulated but no alteration in GSK3 expression profile was observed. Downregulation of NF-κB and upregulation of Caspase 3 and p-β-catenin protein levels were indicated.Conclusion: miR-625-5p can induce apoptosis but it has no significant effect on proliferation of KG-1 cells. MiR-625 can be a promising approach to aid in the treatment of AML. How-ever, further studies are required in this respect.