2026-10-11T19:39:14Z https://api.figshare.com/v2/oai

oai:figshare.com:article/33981424 2026-09-24T10:50:45Z category_7 portal_147 item_type_3 month_year_09_2026

Supplemental Material for: Expression of miR-382-5p in polycystic ovary syndrome and its role in regulating granulosa cell function via targeting TOP1 figshare admin karger (2628495) Min Zhang (111999) Yanqiu Ding (18774130) Yingying Yu (546494) Medicine Medicine <p dir="ltr">Objectives:</p><p dir="ltr">The function of miR-382-5p in polycystic ovary syndrome (PCOS) remains unclear. This study aimed to investigate the potential regulatory mechanisms of the miR-382-5p/DNA topoisomerase I (TOP1) axis in PCOS.</p><p dir="ltr">Design:</p><p dir="ltr">The study included 120 healthy controls and 120 PCOS patients. Serum samples were collected from all participants. Human granulosa-like cell line KGN was cultured in vitro and treated with insulin.</p><p dir="ltr">Methods:</p><p dir="ltr">Reverse transcription quantitative PCR (RT-qPCR) was used to detect RNA expression; the Cell Counting Kit-8 (CCK-8) assay was used to assess cell proliferation; flow cytometry was used to detect cell apoptosis; and enzyme-linked immunosorbent assay (ELISA) was used to measure the protein levels of Bcl-2, Bax, and Cyclin D1. The protein content of TOP1 in KGN cells was detected by Western blotting. RNA immunoprecipitation (RIP) and dual-luciferase assays were used to validate the binding relationship.</p><p dir="ltr">Results:</p><p dir="ltr">Serum miR-382-5p levels were significantly downregulated in PCOS patients, demonstrating good diagnostic value and showing a negative correlation with key clinical indicators of PCOS. In vitro experiments confirmed that, under insulin-induced conditions, overexpression of miR-382-5p significantly inhibited insulin-induced abnormal proliferation of granulosa cells, downregulated the protein of cyclin D1, and simultaneously promoted apoptosis by upregulating the pro-apoptotic protein Bax and downregulating the anti-apoptotic protein Bcl-2. This mechanism was associated with the direct targeting and inhibition of topoisomerase I (TOP1). In PCOS patients, elevated TOP1 mRNA and protein expression correlated negatively with miR-382-5p levels. Overexpression of TOP1 significantly reversed the antiproliferative and proapoptotic effects of miR-382-5p.</p><p dir="ltr">Limitations:</p><p dir="ltr">The verification of the miR 382 5p/TOP1 regulatory mechanism was limited to incomplete in vitro cell experiments and non-standard clinical detection, which cannot fully confirm its exact binding effect, in vivo cascade specificity, and independent pathogenic causality in PCOS patients. Additionally, all PCOS patients presented oligo-/anovulation, whereas healthy controls had regular menstrual cycles; given that circulating miR‑382‑5p levels vary with menstrual cycle phases, the observed downregulation in PCOS serum may partly stem from disrupted cyclic ovulation rather than representing an intrinsic PCOS-specific molecular alteration. Therefore, miR‑382‑5p should be interpreted as a correlative biomarker associated with PCOS phenotypes rather than an independent pathogenic mediator.</p><p dir="ltr">Conclusions:</p><p dir="ltr">miR-382-5p regulated the imbalance between insulin-mediated proliferation and apoptosis in KGN cells by directly targeting TOP1, thereby contributing to the progression of PCOS. </p> 2026-09-24T10:50:45Z Dataset Dataset 10.6084/m9.figshare.33981424.v1 https://figshare.com/articles/dataset/Supplemental_Material_for_Expression_of_miR-382-5p_in_polycystic_ovary_syndrome_and_its_role_in_regulating_granulosa_cell_function_via_targeting_TOP1/33981424 CC BY 4.0