_1788849442 20260908063722000 Rovedar Daryoushbabazadeh@gmail.com Rovedar Journal of Lab Animal Research J Lab Anim. Res 2980-9703 08 30 2026 5 4 Faezeh Barzegar https://orcid.org/0000-0003-1285-0187 Hojjat Ghahvechi https://orcid.org/0009-0003-6312-8626 Gholamhossein Tamaddon https://orcid.org/0000-0001-8158-6004 Zahra Darvish Khalilabadi https://orcid.org/0000-0001-6701-5184 Kamyar Kiamarzi https://orcid.org/0009-0007-1308-015X Mohammad Reza Keramati https://orcid.org/0000-0003-4160-4161 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. 08 30 2026 66 74 https://creativecommons.org/licenses/by/4.0 10.58803/jlar.v5i4.118 https://jlar.rovedar.com/index.php/JLAR/article/view/118 https://jlar.rovedar.com/index.php/JLAR/article/download/118/147 https://jlar.rovedar.com/index.php/JLAR/article/download/118/147 10.1016/S1470-2045(01)00516-2 10.1023/A:1005963732481 10.1038/s41388-017-0091-1 10.1016/S0092-8674(04)00045-5 10.1038/nature02871 10.1073/pnas.0307323101 10.1002/ijc.22454 10.1038/nature03817 10.1038/nature03702 10.1073/pnas.0804549105 10.1038/ncb2037 10.1038/nmeth1079 10.1152/physrev.00027.2001 10.1038/sj.cdd.4401728 10.1089/ars.2013.5371 10.1182/blood-2010-06-288589 10.1038/cddiscovery.2015.8 10.1007/s10495-011-0589-x 10.1016/j.cell.2007.12.018 10.1016/j.febslet.2009.12.034 10.1186/2045-3701-3-35 10.1074/jbc.271.32.19272 10.3892/ol.2017.6509 10.1111/acel.13331 10.5812/semj-143430 10.1038/nprot.2006.83 10.18502/ijpho.v12i3.10058 10.1038/srep36625 10.3390/ani10111951 10.1042/BSR20130098 10.1093/nar/gku376 10.1016/j.tig.2022.02.006 10.1371/journal.pone.0029275 10.18632/oncotarget.4827 10.1016/bs.apha.2017.08.003 10.1093/hmg/ddac256 10.1002/hep.22989 10.1038/leu.2013.361 10.1080/15548627.2016.1245263 10.1016/j.mcp.2015.08.002 10.4161/auto.23117