Tabriz University of Medical Sciences Advanced Pharmaceutical Bulletin 2228-5881 15 3 2025 09 25 Continuous Uptake of MiR-181a-2 Mimetic Induces Constitutive Overexpression of Cellular MiR-181a-2 in MCF-7 Breast Cancer Cells: Links with Progression of Drug Resistance 678 685 10.34172/apb.025.43440 EN Olga Evgen’evna Andreeva https://orcid.org/0000-0002-6015-6619 Danila Vladimirovich Sorokin https://orcid.org/0000-0002-1264-7405 Svetlana Vladimirovna Vinokurova https://orcid.org/0000-0003-1615-3928 Pavel Borisovich Kopnin https://orcid.org/0000-0002-2078-4274 Nadezhda Viacheslavovna Elkina https://orcid.org/0000-0002-0503-6016 Danila Sergeevich Elkin https://orcid.org/0000-0002-4793-6063 Maria Dmitrievna Fedorova https://orcid.org/0000-0002-8813-7516 Alexander Mikhailovich Scherbakov https://orcid.org/0000-0002-2974-9555 Mikhail Aleksandrovich Krasil’nikov https://orcid.org/0000-0002-5902-7633 Journal Article 10.34172/apb.025.43440 2024 07 28 2025 08 25 Purpose: The aim of this study was to elucidate the mechanisms underlying the formation and maintenance of drug resistance in cancer cells. Previously, we demonstrated that prolonged treatment of estrogen-dependent MCF-7 breast cancer cells with exosomes derived from estrogen-resistant MCF-7/T cells leads to a partial loss of estrogen sensitivity in MCF-7 cells. Moreover, repeated transfection with one of the exosomal microRNAs—microRNA-181a-2—induced an irreversible decrease in hormonal sensitivity in the recipient cells. In the present work, to further investigate the possible mechanism of miR-181a-2-induced acquired resistance, we analyzed the effect of multiple miR-181a-2 transfections on the expression of cellular miR-181a-2 and related signaling proteins. Methods: miR-181a-2 was ectopically expressed by mimetic transfection or suppressed by antisense oligonucleotides. miR-181a-2 precursor/MIR181A2HG expression (qRT-PCR) and MIR181A2 locus copy number (qPCR) were assessed. wtSnail was expressed via transient transfection. Tamoxifen sensitivity was measured by MTT assay. Protein expression was studied by immunoblotting, estrogen receptor α/Snail transcriptional activity was evaluated by reporter analysis. Results: We found that multiple transfections with miR-181a-2 resulted in a marked increase in cellular miR-181a-2 precursor levels, whereas single transfection had no such effect. Similarly, stable transfection with miR-181a-2 led to increased levels of cellular miR-181a-2 and its host gene, MIR181A2HG, which was associated with partial resistance to tamoxifen. Analysis of the genomic DNA encoding miR-181a-2 revealed no changes in copy number in transfected cells. Furthermore, we identified the transcription factor Snail as a key mediator of miR-181a-2–induced resistance and demonstrated its role in the formation of an autoregulatory loop of miR181a-2 and the maintenance of cell resistance. Conclusion: Overall, these results reveal a novel mechanism of resistance-associated signaling pathway rearrangement based on the formation of a miR-181a-2 autoregulatory loop.