{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,27]],"date-time":"2026-07-27T09:07:40Z","timestamp":1785143260739,"version":"3.55.0"},"reference-count":38,"publisher":"PeerJ","license":[{"start":{"date-parts":[[2026,7,27]],"date-time":"2026-07-27T00:00:00Z","timestamp":1785110400000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Middle\/Young aged Teachers\u2019 Research Ability Improvement Project of Guangxi Higher Education","award":["No. 2024KY0119"],"award-info":[{"award-number":["No. 2024KY0119"]}]},{"name":"Guangxi Natural Science Foundation","award":["No. 2025GXNSFAA069816"],"award-info":[{"award-number":["No. 2025GXNSFAA069816"]}]},{"name":"Scientific Research Project of Liuzhou People\u2019s Hospital affiliated to Guangxi Medical University","award":["No. lry202327"],"award-info":[{"award-number":["No. lry202327"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Diminished ovarian reserve (DOR) is a common cause of female infertility. Prior high-throughput sequencing showed specific follicular fluid exosomal miRNA profiles in DOR patients, implicating exosomal miRNAs in DOR pathogenesis. Using miRNA PCR arrays, we found miR-483-3p was significantly upregulated in DOR follicular fluid exosomes. Bioinformatics and experiments indicated METTL3 as a potential miR-483-3p target, suggesting excess miR-483-3p exerts cytotoxicity and suppresses METTL3-mediated N6-methyladenosine (m6A) modification. This study thus investigated miR-483-3p effects on METTL3 expression, m6A levels, cell proliferation, and mitochondrial function in ovarian granulosa cells (GCs).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>The targeting interaction between miR-483-3p and METTL3 was confirmed using a dual-luciferase reporter gene assay. qRT-PCR, immunofluorescence, Western blot, and Dot blot were used to evaluate the effects of miR-483-3p mimics, cyclophosphamide (CTX), and STM2457 on METTL3 protein expression and m6A modification levels in KGN cells. Additionally, Cell Counting Kit-8 (CCK-8) and 5-Ethynyl-2\u2019-deoxyuridine (EdU) assays, along with mitochondrial membrane potential (MMP) measurements, were used to examine the impact of these treatments on KGN cell proliferation and mitochondrial function.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>miR-483-3p directly targeted the 3\u2032\u00a0untranslated region (3\u2032UTR) of METTL3, and its mimics significantly suppressed METTL3 gene expression. In contrast, CTX robustly promoted METTL3 gene expression, accompanied by a substantial elevation in global m6A levels. STM2457 treatment also showed a similar trend. But miR-483-3p overexpression modestly inhibited METTL3 protein and m6A levels. Regarding cellular phenotypes, miR-483-3p overexpression, CTX, and STM2457 all exerted significant inhibitory effects on KGN cells. All three treatments consistently suppressed the cell viability of KGN cells, reduced the EdU-positive cell ratio, and decreased MMP levels.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      This study provides preliminary evidence suggesting a specific regulatory interaction between miR-483-3p and METTL3. We demonstrate that aberrantly high expression of miR-483-3p significantly suppresses METTL3 mRNA levels but only moderately reduces METTL3 protein and m6A levels, suggesting the involvement of complex post-transcriptional regulatory mechanisms. Furthermore, overaccumulation of miR-483-3p markedly inhibits the proliferation of ovarian GCs and impairs their mitochondrial function. Further investigation into the role of the candidate miR-483-3p-METTL3-m6A axis in DOR will be conducted using\n                      <jats:italic>in vivo<\/jats:italic>\n                      models alongside an expanded cohort of clinical samples.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.7717\/peerj.21567","type":"journal-article","created":{"date-parts":[[2026,7,27]],"date-time":"2026-07-27T08:27:00Z","timestamp":1785140820000},"page":"e21567","source":"Crossref","is-referenced-by-count":0,"title":["Overaccumulation of miR-483-3p exerts acute toxic effects on ovarian granulosa cells by impairing cell proliferation, mitochondrial function, and METTL3-mediated m6A modification"],"prefix":"10.7717","volume":"14","author":[{"given":"Kaiyuan","family":"Shen","sequence":"first","affiliation":[{"name":"Reproductive Medicine, Liuzhou People\u2019s Hospital Affiliated to Guangxi Medical University, Liuzhou,  Guangxi, China"},{"name":"Liuzhou Key Laboratory of Reproductive and Genetic Metabolic Diseases, Liuzhou, Guangxi, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoli","family":"Dai","sequence":"additional","affiliation":[{"name":"Research Service Office, Liuzhou People\u2019s Hospital Affiliated to Guangxi Medical University, Liuzhou,  Guangxi, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liqun","family":"Chen","sequence":"additional","affiliation":[{"name":"Reproductive Medicine, Liuzhou People\u2019s Hospital Affiliated to Guangxi Medical University, Liuzhou,  Guangxi, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuanyuan","family":"Liang","sequence":"additional","affiliation":[{"name":"Reproductive Medicine, Liuzhou People\u2019s Hospital Affiliated to Guangxi Medical University, Liuzhou,  Guangxi, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ping","family":"Huang","sequence":"additional","affiliation":[{"name":"Reproductive Medicine, Liuzhou People\u2019s Hospital Affiliated to Guangxi Medical University, Liuzhou,  Guangxi, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuxin","family":"Zeng","sequence":"additional","affiliation":[{"name":"Reproductive Medicine, Liuzhou People\u2019s Hospital Affiliated to Guangxi Medical University, Liuzhou,  Guangxi, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoli","family":"Qu","sequence":"additional","affiliation":[{"name":"Reproductive Medicine, Liuzhou People\u2019s Hospital Affiliated to Guangxi Medical University, Liuzhou,  Guangxi, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"4443","published-online":{"date-parts":[[2026,7,27]]},"reference":[{"issue":"16","key":"10.7717\/peerj.21567\/ref-1","doi-asserted-by":"publisher","first-page":"5834","DOI":"10.3390\/jcm14165834","article-title":"The final phases of ovarian aging: a tale of diverging functional trajectories","volume":"14","author":"Bochynska","year":"2025","journal-title":"Journal of Clinical Medicine"},{"issue":"1","key":"10.7717\/peerj.21567\/ref-2","doi-asserted-by":"publisher","first-page":"61","DOI":"10.1186\/s12964-022-00876-6","article-title":"Follicular fluid-derived exosomal miR-143-3p\/miR-155-5p regulate follicular dysplasia by modulating glycolysis in granulosa cells in polycystic ovary syndrome","volume":"20","author":"Cao","year":"2022","journal-title":"Cell Communication and Signaling"},{"issue":"1","key":"10.7717\/peerj.21567\/ref-3","doi-asserted-by":"publisher","first-page":"107","DOI":"10.1186\/s13048-024-01434-z","article-title":"Cyclophosphamide induces ovarian granulosa cell ferroptosis via a mechanism associated with HO-1 and ROS-mediated mitochondrial dysfunction","volume":"17","author":"Chen","year":"2024","journal-title":"Journal of Ovarian Research"},{"issue":"3","key":"10.7717\/peerj.21567\/ref-4","doi-asserted-by":"publisher","first-page":"925","DOI":"10.14245\/ns.2448494.247","article-title":"STM2457 inhibits METTL3-mediated m6A modification of miR-30c to alleviate spinal cord injury by inducing the ATG5-mediated autophagy","volume":"21","author":"Chen","year":"2024","journal-title":"Neurospine"},{"issue":"1","key":"10.7717\/peerj.21567\/ref-5","doi-asserted-by":"publisher","first-page":"7","DOI":"10.1038\/s41389-020-00290-y","article-title":"Metformin exhibits antiproliferation activity in breast cancer via miR-483-3p\/METTL3\/m(6)A\/p21 pathway","volume":"10","author":"Cheng","year":"2021","journal-title":"Oncogenesis"},{"key":"10.7717\/peerj.21567\/ref-6","doi-asserted-by":"publisher","first-page":"156224","DOI":"10.1016\/j.prp.2025.156224","article-title":"METTL3 promotes ovarian cancer progression through YTHDF2-dependent degradation of GATA4","volume":"275","author":"Di","year":"2025","journal-title":"Pathology, Research and Practice"},{"issue":"3","key":"10.7717\/peerj.21567\/ref-7","doi-asserted-by":"publisher","first-page":"303","DOI":"10.1007\/s10815-016-0657-9","article-title":"Exosome-mediated communication in the ovarian follicle","volume":"33","author":"Di\u00a0Pietro","year":"2016","journal-title":"Journal of Assisted Reproduction and Genetics"},{"issue":"4","key":"10.7717\/peerj.21567\/ref-8","doi-asserted-by":"publisher","first-page":"2165","DOI":"10.3390\/ijms22042165","article-title":"Exosomes as biomarkers for female reproductive diseases diagnosis and therapy","volume":"22","author":"Esfandyari","year":"2021","journal-title":"International Journal of Molecular Sciences"},{"key":"10.7717\/peerj.21567\/ref-9","doi-asserted-by":"publisher","first-page":"1072313","DOI":"10.3389\/fendo.2022.1072313","article-title":"Regulation of ovarian function by growth hormone: Potential intervention of ovarian aging","volume":"13","author":"Han","year":"2022","journal-title":"Frontiers in Endocrinology"},{"key":"10.7717\/peerj.21567\/ref-10","doi-asserted-by":"publisher","first-page":"415","DOI":"10.3389\/fendo.2019.00415","article-title":"Cyclophosphamide regulates N6-Methyladenosine and m6A RNA enzyme levels in human granulosa cells and in ovaries of a premature ovarian aging mouse model","volume":"10","author":"Huang","year":"2019","journal-title":"Frontiers in Endocrinology"},{"issue":"1","key":"10.7717\/peerj.21567\/ref-11","doi-asserted-by":"publisher","first-page":"693","DOI":"10.1038\/s41598-024-84437-6","article-title":"hsa-mir-483-3p modulates delayed breast cancer recurrence","volume":"15","author":"Kim","year":"2025","journal-title":"Scientific Reports"},{"key":"10.7717\/peerj.21567\/ref-12","doi-asserted-by":"publisher","first-page":"116820","DOI":"10.1016\/j.bcp.2025.116820","article-title":"STM2457 decreases m6A methylation to reduce cisplatin-induced ototoxicity via MAPK signaling","volume":"235","author":"Liao","year":"2025","journal-title":"Biochemical Pharmacology"},{"issue":"11","key":"10.7717\/peerj.21567\/ref-13","doi-asserted-by":"publisher","first-page":"9205","DOI":"10.3390\/ijms24119205","article-title":"Mechanisms of and potential medications for oxidative stress in ovarian granulosa cells: a review","volume":"24","author":"Liu","year":"2023","journal-title":"International Journal of Molecular Sciences"},{"issue":"1","key":"10.7717\/peerj.21567\/ref-14","doi-asserted-by":"publisher","first-page":"160","DOI":"10.1186\/s12958-022-01037-7","article-title":"hsa-miR-320a-3p and hsa-miR-483-5p levels in human granulosa cells: promising bio-markers of live birth after IVF\/ICSI","volume":"20","author":"Liu","year":"2022","journal-title":"Reproductive Biology and Endocrinology"},{"issue":"1","key":"10.7717\/peerj.21567\/ref-15","doi-asserted-by":"publisher","first-page":"320","DOI":"10.1186\/s13287-022-03012-w","article-title":"Cell-free fat extract improves ovarian function and fertility in mice with premature ovarian insufficiency","volume":"13","author":"Liu","year":"2022","journal-title":"Stem Cell Research & Therapy"},{"key":"10.7717\/peerj.21567\/ref-16","doi-asserted-by":"publisher","first-page":"144","DOI":"10.3389\/fmicb.2020.00144","article-title":"High levels of miR-483-3p are present in serum exosomes upon infection of mice with highly pathogenic avian influenza virus","volume":"11","author":"Maemura","year":"2020","journal-title":"Frontiers in Microbiology"},{"issue":"9","key":"10.7717\/peerj.21567\/ref-17","doi-asserted-by":"publisher","first-page":"1372","DOI":"10.1093\/infdis\/jiy035","article-title":"Lung-derived exosomal miR-483-3p regulates the innate immune response to influenza virus infection","volume":"217","author":"Maemura","year":"2018","journal-title":"The Journal of Infectious Diseases"},{"issue":"11","key":"10.7717\/peerj.21567\/ref-18","doi-asserted-by":"publisher","first-page":"989","DOI":"10.1038\/s41419-021-04272-9","article-title":"METTL3-mediated mRNA N(6)-methyladenosine is required for oocyte and follicle development in mice","volume":"12","author":"Mu","year":"2021","journal-title":"Cell Death & Disease"},{"issue":"1","key":"10.7717\/peerj.21567\/ref-19","doi-asserted-by":"publisher","first-page":"17","DOI":"10.1007\/s10815-017-1058-4","article-title":"Reproductive ovarian testing and the alphabet soup of diagnoses: DOR, POI, POF, POR, and FOR","volume":"35","author":"Pastore","year":"2018","journal-title":"Journal of Assisted Reproduction and Genetics"},{"key":"10.7717\/peerj.21567\/ref-20","doi-asserted-by":"publisher","first-page":"814240","DOI":"10.3389\/fmolb.2022.814240","article-title":"Diagnostic potential of plasma extracellular vesicle miR-483-3p and Let-7d-3p for sepsis","volume":"9","author":"Qiu","year":"2022","journal-title":"Frontiers in Molecular Biosciences"},{"issue":"17","key":"10.7717\/peerj.21567\/ref-21","doi-asserted-by":"publisher","first-page":"5105","DOI":"10.3390\/jcm13175105","article-title":"Metabolic risk in patients with a diminished ovarian reserve and premature ovarian insufficiency","volume":"13","author":"Robeva","year":"2024","journal-title":"Journal of Clinical Medicine"},{"issue":"5","key":"10.7717\/peerj.21567\/ref-22","doi-asserted-by":"publisher","first-page":"916","DOI":"10.1093\/biolre\/ioaf034","article-title":"Age-related integrative transcriptomic profiling of human granulosa cells reveals mRNA-microRNA regulatory network associated with key ovulation dynamics\u2020","volume":"112","author":"Rogers","year":"2025","journal-title":"Biology of Reproduction"},{"issue":"6","key":"10.7717\/peerj.21567\/ref-23","doi-asserted-by":"publisher","first-page":"1751","DOI":"10.1016\/j.fertnstert.2014.08.005","article-title":"Molecular characterization of exosomes and their microRNA cargo in human follicular fluid: bioinformatic analysis reveals that exosomal microRNAs control pathways involved in follicular maturation","volume":"102","author":"Santonocito","year":"2014","journal-title":"Fertility and Sterility"},{"key":"10.7717\/peerj.21567\/ref-24","doi-asserted-by":"publisher","first-page":"2163","DOI":"10.2147\/ott.S288936","article-title":"Upregulated MicroRNA-483-3p is an early event in pancreatic ductal adenocarcinoma (PDAC) and as a powerful liquid biopsy biomarker in PDAC","volume":"14","author":"Shao","year":"2021","journal-title":"OncoTargets and Therapy"},{"issue":"1","key":"10.7717\/peerj.21567\/ref-25","doi-asserted-by":"publisher","first-page":"308","DOI":"10.1186\/s12920-023-01756-9","article-title":"Specific expression profile of follicular fluid-derived exosomal microRNAs in patients with diminished ovarian reserve","volume":"16","author":"Shen","year":"2023","journal-title":"BMC Medical Genomics"},{"issue":"2","key":"10.7717\/peerj.21567\/ref-26","doi-asserted-by":"publisher","first-page":"174","DOI":"10.1071\/rd21277","article-title":"Current knowledge and the future potential of extracellular vesicles in mammalian reproduction","volume":"34","author":"Tesfaye","year":"2021","journal-title":"Reproduction, Fertility, and Development"},{"key":"10.7717\/peerj.21567\/ref-27","doi-asserted-by":"publisher","first-page":"108901","DOI":"10.1016\/j.reprotox.2025.108901","article-title":"Exposure to disinfection by-products and risk of diminished ovarian reserve: case-control evidence and cellular metabolomic insights","volume":"135","author":"Tian","year":"2025","journal-title":"Reproductive Toxicology"},{"key":"10.7717\/peerj.21567\/ref-28","doi-asserted-by":"publisher","first-page":"110519","DOI":"10.1016\/j.biopha.2020.110519","article-title":"miR-483-3p promotes cell proliferation and suppresses apoptosis in rheumatoid arthritis fibroblast-like synoviocytes by targeting IGF-1","volume":"130","author":"Wang","year":"2020","journal-title":"Biomedicine & Pharmacotherapy"},{"issue":"9","key":"10.7717\/peerj.21567\/ref-29","doi-asserted-by":"publisher","first-page":"1194","DOI":"10.1038\/s43587-024-00697-x","article-title":"Rejuvenation of aged oocyte through exposure to young follicular microenvironment","volume":"4","author":"Wang","year":"2024","journal-title":"Nature Aging"},{"key":"10.7717\/peerj.21567\/ref-30","doi-asserted-by":"publisher","first-page":"216122","DOI":"10.1016\/j.canlet.2023.216122","article-title":"METTL3-m(6)A-EGFR-axis drives lenvatinib resistance in hepatocellular carcinoma","volume":"559","author":"Wang","year":"2023","journal-title":"Cancer Letters"},{"issue":"7860","key":"10.7717\/peerj.21567\/ref-31","doi-asserted-by":"publisher","first-page":"597","DOI":"10.1038\/s41586-021-03536-w","article-title":"Small-molecule inhibition of METTL3 as a strategy against myeloid leukaemia","volume":"593","author":"Yankova","year":"2021","journal-title":"Nature"},{"issue":"5","key":"10.7717\/peerj.21567\/ref-32","doi-asserted-by":"publisher","first-page":"340","DOI":"10.1038\/s41419-023-05859-0","article-title":"Chemotherapy impairs ovarian function through excessive ROS-induced ferroptosis","volume":"14","author":"Zhang","year":"2023","journal-title":"Cell Death & Disease"},{"key":"10.7717\/peerj.21567\/ref-33","doi-asserted-by":"publisher","first-page":"468","DOI":"10.1016\/j.bbrc.2020.11.058","article-title":"Association of exosomal microRNAs in human ovarian follicular fluid with oocyte quality","volume":"534","author":"Zhang","year":"2021","journal-title":"Biochemical and Biophysical Research Communications"},{"key":"10.7717\/peerj.21567\/ref-34","doi-asserted-by":"publisher","first-page":"1477665","DOI":"10.3389\/fendo.2024.1477665","article-title":"Subclinical\/overt hypothyroidism may be associated with diminished ovarian reserve in infertile women independent of thyroid autoimmunity","volume":"15","author":"Zhang","year":"2024","journal-title":"Frontiers in Endocrinology"},{"issue":"1","key":"10.7717\/peerj.21567\/ref-35","doi-asserted-by":"publisher","first-page":"4359","DOI":"10.1038\/s41598-022-08423-6","article-title":"Exosomal miR-143-3p derived from follicular fluid promotes granulosa cell apoptosis by targeting BMPR1A in polycystic ovary syndrome","volume":"12","author":"Zhao","year":"2022","journal-title":"Scientific Reports"},{"issue":"1","key":"10.7717\/peerj.21567\/ref-36","doi-asserted-by":"publisher","first-page":"20","DOI":"10.1002\/jcp.31140","article-title":"miRNA profiling of granulosa cell-derived exosomes reveals their role in promoting follicle development","volume":"239","author":"Zhou","year":"2024","journal-title":"Journal of Cellular Physiology"},{"issue":"4","key":"10.7717\/peerj.21567\/ref-37","doi-asserted-by":"publisher","first-page":"2520","DOI":"10.1007\/s12035-021-02714-1","article-title":"Follicular fluid-derived exosomal MicroRNA-18b-5p regulates PTEN-mediated PI3K\/Akt\/mTOR signaling pathway to inhibit polycystic ovary syndrome development","volume":"59","author":"Zhou","year":"2022","journal-title":"Molecular Neurobiology"},{"issue":"1","key":"10.7717\/peerj.21567\/ref-38","doi-asserted-by":"publisher","first-page":"208","DOI":"10.1186\/s13048-023-01296-x","article-title":"Potential factors result in diminished ovarian reserve: a comprehensive review","volume":"16","author":"Zhu","year":"2023","journal-title":"Journal of Ovarian Research"}],"container-title":["PeerJ"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/peerj.com\/articles\/21567.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/peerj.com\/articles\/21567.xml","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/peerj.com\/articles\/21567.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/peerj.com\/articles\/21567.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,27]],"date-time":"2026-07-27T08:27:04Z","timestamp":1785140824000},"score":1,"resource":{"primary":{"URL":"https:\/\/peerj.com\/articles\/21567"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,27]]},"references-count":38,"alternative-id":["10.7717\/peerj.21567"],"URL":"https:\/\/doi.org\/10.7717\/peerj.21567","archive":["CLOCKSS","LOCKSS","Portico"],"relation":{},"ISSN":["2167-8359"],"issn-type":[{"value":"2167-8359","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,27]]},"article-number":"e21567"}}