In Silico and In Vitro Analysis of the Regulatory Role of miR-145-5p on Its Potential Target Genes in Acute Myeloid Leukemia Cell Lines

Abstract

Objective: Acute myeloid leukemia (AML) demonstrates prognostic heterogeneity, and the underlying pathophysiology is still not fully elucidated. This study investigated how the tumor suppressor miR-145-5p contributes to AML heterogeneity and associated biological processes through its potential target onco genes. Materials and Methods: HL-60 and NB4 AML cells were transfected with miR-145-5p mimics using Lipo fectamine™ 2000. Apoptosis was evaluated by caspase-3 activity in NB4 cells, while cell viability was determined using WST-8 assays in HL-60 and NB4 cells. Putative miR-145-5p targets were predicted using the miRTarBase (v9.0) and miRNET (v2.0) databases. Following further filtration, PPI, KEGG, and GO analyses were performed using in silico approaches. Expression profiling of the identified genes was evaluated via quantitative real-time polymerase chain reaction (qRT-PCR), and the data were statistically compared. Results: After miR-145-5p transfection, cell survival markedly decreased in both cell lines, and caspase-3 levels markedly increased in NB4 cells (p<0.05). Among the putative genes CDK4, CDK6, KLF4, NRAS, IRS1, and eIF4E identified via in silico analysis, CDK4 and KLF4 showed significantly decreased expression in both cell lines (p<0.05 for both). Conclusion: The results demonstrate that miR-145-5p potentially regulates apoptosis and proliferation in AML via target genes, including CDK4 and KLF4. Further studies may reinforce the biomarker potential of miR-145-5p and facilitate the advancement of innovative diagnostic and therapeutic approaches for AML.

Keywords

• AML
• miR-145-5p
• HL-60
• NB4
• CDK4
• KLF4

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