eng Rovedar Journal of Lab Animal Research 2980-9703 2026-08-30 5 4 66 74 10.58803/jlar.v5i4.118 128 Faezeh Barzegar 0 https://orcid.org/0000-0003-1285-0187 Hojjat Ghahvechi 0 https://orcid.org/0009-0003-6312-8626 Gholamhossein Tamaddon 1 https://orcid.org/0000-0001-8158-6004 Zahra Darvish Khalilabadi 2 https://orcid.org/0000-0001-6701-5184 Kamyar Kiamarzi 3 https://orcid.org/0009-0007-1308-015X Mohammad Reza Keramati 4 https://orcid.org/0000-0003-4160-4161 Department of Hematology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran Department of Hematology, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran Department of Hematology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran. Cancer Molecular Pathology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran Student Research Committee, Jahrom University of Medical Sciences, Jahrom, Iran Cancer Molecular Pathology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran Introduction: MicroRNAs (miRNAs) can affect cellular processes by modulating the expression of different genes. Blocking specific miRNAs across all cancer types can reduce or prevent cancer cell activity. The present study aimed to investigate the inhibitory effects of MicroRNA sponges (miRSponge) on miR-17-5p and miR-181 in NALM-6 cells, a cellular model of acute lymphoblastic leukemia, and to evaluate their effects on the expression of the autophagy-related genes ATG-5 and ATG-7. Materials and methods: The present study was conducted from 2021 to 2022 at Mashhad University of Medical Sciences, Iran, in collaboration with the Shiraz Paramedical Faculty, Iran, and focused on a miRSponge designed to target miR-17-5p and miR-181. The present study included four NALM-6 cell groups, including untreated cells as a control, miR-17-5p- and miR-181-sponge-transfected cells, and a vincristine-treated group. The NALM-6 cells were cultured and transfected with a miRSponge targeting miR-17-5p and miR-181. The expression levels of miR-17-5p, miR-181, ATG-5, and ATG-7 were quantified by real-time quantitative PCR, while apoptosis in cells from different study groups was assessed by flow cytometry using Annexin V/7-AAD staining. Cell viability following vincristine treatment was evaluated using MTT assay, and bioinformatics analyses were performed to investigate miRNA-target interactions. Results: The current results indicated that the developed sponge significantly reduced the levels of these two oncogenic miRNAs in cells by binding to miR-17-5p and miR-181. Furthermore, the activity of ATG-5 and ATG-7, regulated by these two miRNAs, increased following the reduction in the expression of these miRNAs. Late-stage apoptosis was higher in miRSponge-treated cells than in vincristine-treated and control groups, although the difference was not statistically significant. Conclusion: The present findings indicated that using a miRSponge significantly upregulated the ATG-5 and ATG-7 genes and enhanced autophagy and apoptosis in cancer cells. MicroRNA sponges reduced the effects of oncogenic miRNAs in acute lymphocytic leukemia cells. https://jlar.rovedar.com/index.php/JLAR/article/view/118 Acute lymphocytic leukemia miRNA miRSponge miR-17-5P miR-181