{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T15:59:29Z","timestamp":1785427169470,"version":"3.56.0"},"reference-count":27,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2026,7,23]],"date-time":"2026-07-23T00:00:00Z","timestamp":1784764800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004085","name":"Soonchunhyang University Research Fund","doi-asserted-by":"publisher","award":["RS-2024-00333544"],"award-info":[{"award-number":["RS-2024-00333544"]}],"id":[{"id":"10.13039\/501100004085","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003725","name":"National Research Foundation of Korea","doi-asserted-by":"crossref","award":["RS-2023-00219563"],"award-info":[{"award-number":["RS-2023-00219563"]}],"id":[{"id":"10.13039\/501100003725","id-type":"DOI","asserted-by":"crossref"}]},{"award":["RS-2023-00219563"],"award-info":[{"award-number":["RS-2023-00219563"]}],"id":[{"id":"https:\/\/ror.org\/013aysd81","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceuticals"],"abstract":"<jats:p>Background\/Objectives: Prostate cancer (PCa) remains a leading cause of cancer-related mortality in men. While localized PCa carries a favorable prognosis, progression to locally advanced disease substantially worsens outcomes. There is an unmet need for molecular markers that reliably distinguish indolent from aggressive tumors. MicroRNAs (miRNAs), small non-coding RNAs that post-transcriptionally regulate gene expression, are implicated in cancer development and progression, and their expression profiles may serve as putative biomarkers of disease stage. Methods: We performed small RNA sequencing on formalin-fixed paraffin-embedded (FFPE) prostate tumor tissues from nine patients (five localized, four locally advanced-stage). Differentially expressed miRNAs were identified using a bioinformatics pipeline (fold-change \u2265 2, p &lt; 0.05). Target gene prediction, KEGG pathway enrichment, and miRNA\u2013mRNA interaction network analyses were conducted, followed by in silico validation using two independent public mRNA datasets (GSE46602 and GSE21034, a subseries of GSE21032). Results: Seven unique mature miRNAs were differentially expressed between localized and locally advanced PCa. Five miRNAs (miR-145-5p, miR-205-5p, miR-206, miR-24-1-5p, and hsa-miR-3648) were downregulated in locally advanced tumors, while hsa-miR-625-3p and miR-324-5p were upregulated. Network analysis identified CLTC and CDK6 as putative downstream targets co-targeted by multiple downregulated miRNAs. Independent validation using GSE46602 and GSE21034 consistently demonstrated significantly higher expression of CLTC and CDK6 in locally advanced prostate cancer following Benjamini\u2013Hochberg correction. Conclusions: This exploratory pilot analysis identified several miRNAs that differ between localized (pT2) and locally advanced (pT3\u2013T4\/N+) prostate cancer. Independent validation across two publicly available mRNA cohorts supports CLTC and CDK6 as candidate stage-associated genes. However, these findings remain hypothesis-generating and require validation in larger prospective cohorts together with functional confirmation before biomarker or therapeutic claims can be made.<\/jats:p>","DOI":"10.3390\/ph19081141","type":"journal-article","created":{"date-parts":[[2026,7,23]],"date-time":"2026-07-23T15:05:42Z","timestamp":1784819142000},"page":"1141","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Exploratory Small RNA Sequencing in Localized Versus Locally Advanced Prostate Cancer: Putative Roles of CLTC and CDK6 as Downstream Targets of Differentially Expressed MicroRNAs"],"prefix":"10.3390","volume":"19","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4490-3610","authenticated-orcid":false,"given":"Jae Heon","family":"Kim","sequence":"first","affiliation":[{"name":"Department of Urology, Soonchunhyang University School of Medicine, Seoul 04404, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ahrim","family":"Moon","sequence":"additional","affiliation":[{"name":"Department of Pathology, Soonchunhyang University School of Medicine, Bucheon 14584, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-8345-6686","authenticated-orcid":false,"given":"Miho","family":"Song","sequence":"additional","affiliation":[{"name":"Department of Urology, Soonchunhyang University School of Medicine, Seoul 04404, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8882-2492","authenticated-orcid":false,"given":"Kwang Woo","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Urology, Soonchunhyang University School of Medicine, Bucheon 14584, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3031-1120","authenticated-orcid":false,"given":"Soo Min","family":"Suh","sequence":"additional","affiliation":[{"name":"Soonchunhyang Institute of Medi-Bio Science (SIMS), Soonchunhyang University, Cheonan 31151, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-3431-2157","authenticated-orcid":false,"given":"Hui Ji","family":"Kim","sequence":"additional","affiliation":[{"name":"Soonchunhyang Institute of Medi-Bio Science (SIMS), Soonchunhyang University, Cheonan 31151, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luis Alfonso","family":"Pefianco","sequence":"additional","affiliation":[{"name":"Soonchunhyang Institute of Medi-Bio Science (SIMS), Soonchunhyang University, Cheonan 31151, Republic of Korea"},{"name":"Department of Integrated Biomedical Science, Soonchunhyang University, Cheonan 31151, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kevin","family":"Andrean","sequence":"additional","affiliation":[{"name":"Soonchunhyang Institute of Medi-Bio Science (SIMS), Soonchunhyang University, Cheonan 31151, Republic of Korea"},{"name":"Department of Integrated Biomedical Science, Soonchunhyang University, Cheonan 31151, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0825-3996","authenticated-orcid":false,"given":"Kisoo","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Urology, Dong-A University College of Medicine, Busan 49201, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seongho","family":"Ryu","sequence":"additional","affiliation":[{"name":"Soonchunhyang Institute of Medi-Bio Science (SIMS), Soonchunhyang University, Cheonan 31151, Republic of Korea"},{"name":"Department of Integrated Biomedical Science, Soonchunhyang University, Cheonan 31151, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yun-Seob","family":"Song","sequence":"additional","affiliation":[{"name":"Department of Urology, Soonchunhyang University School of Medicine, Seoul 04404, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,23]]},"reference":[{"key":"ref_1","first-page":"209","article-title":"Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries","volume":"71","author":"Sung","year":"2021","journal-title":"CA Cancer J. Clin."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"8165","DOI":"10.1200\/JCO.2005.03.3134","article-title":"Active surveillance for prostate cancer: For whom?","volume":"23","author":"Klotz","year":"2005","journal-title":"J. Clin. Oncol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"3576","DOI":"10.1200\/JCO.2006.10.3820","article-title":"A nomogram predicting 10-year life expectancy in candidates for radical prostatectomy or radiotherapy for prostate cancer","volume":"25","author":"Walz","year":"2007","journal-title":"J. Clin. Oncol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"878","DOI":"10.1093\/jnci\/djp122","article-title":"Risk Assessment for Prostate Cancer Metastasis and Mortality at the Time of Diagnosis","volume":"101","author":"Cooperberg","year":"2009","journal-title":"JNCI J. Natl. Cancer Inst."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"439","DOI":"10.1002\/jcb.20522","article-title":"A paradigm for the treatment of prostate cancer bone metastases based on an understanding of tumor cell\u2013microenvironment interactions","volume":"96","author":"Loberg","year":"2005","journal-title":"J. Cell. Biochem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"7771","DOI":"10.1038\/s41598-025-92091-9","article-title":"MiRNA expression profiling and clinical implications in prostate cancer across various stages","volume":"15","author":"Xiang","year":"2025","journal-title":"Sci. Rep."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"J\u00e4remo, H., Semenas, J., Bergstr\u00f6m, S.H., Lundholm, M., Thysell, E., Widmark, A., Crnalic, S., Ylitalo, E.B., Bergh, A., and Brattsand, M. (2023). Investigating microRNA profiles in prostate cancer bone metastases and functional effects of microRNA-23c and microRNA-4328. Cancers, 15.","DOI":"10.3390\/cancers15092437"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3646","DOI":"10.1038\/s41598-025-85502-4","article-title":"Comprehensive computational analysis of differentially expressed miRNAs and their influence on transcriptomic signatures in prostate cancer","volume":"15","author":"Aldakheel","year":"2025","journal-title":"Sci. Rep."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"13864","DOI":"10.1038\/s41598-021-93208-6","article-title":"High expression of miR-17-5p in tumor epithelium is a predictor for poor prognosis for prostate cancer patients","volume":"11","author":"Stoen","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"e16705","DOI":"10.1097\/MD.0000000000016705","article-title":"Increased expression of miR-153 predicts poor prognosis for patients with prostate cancer","volume":"98","author":"Bi","year":"2019","journal-title":"Medicine"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"479","DOI":"10.1002\/pros.23468","article-title":"MiR-206 inhibits proliferation and migration of prostate cancer cells by targeting CXCL11","volume":"78","author":"Wang","year":"2018","journal-title":"Prostate"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1038\/s10038-017-0371-1","article-title":"Regulation of HMGB3 by antitumor miR-205-5p inhibits cancer cell aggressiveness","volume":"63","author":"Yamada","year":"2018","journal-title":"J. Hum. Genet."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"707","DOI":"10.1038\/bjc.2016.233","article-title":"The utility of urine-circulating miRNAs for detection of prostate cancer","volume":"115","author":"Daniunaite","year":"2016","journal-title":"Br. J. Cancer"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1186\/s12967-019-1920-5","article-title":"Circulating miRNAs as non-invasive biomarkers to predict aggressive prostate cancer after radical prostatectomy","volume":"17","author":"Hoey","year":"2019","journal-title":"J. Transl. Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"768","DOI":"10.1038\/bjc.2011.595","article-title":"Changes in circulating microRNA levels associated with prostate cancer","volume":"106","author":"Bryant","year":"2012","journal-title":"Br. J. Cancer"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"4412705","DOI":"10.1155\/2021\/4412705","article-title":"MiR-145-5p inhibits prostate cancer invasion and induces apoptosis by Inhibiting WIP1","volume":"2021","author":"Sun","year":"2021","journal-title":"J. Oncol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1016\/j.prnil.2014.09.001","article-title":"Prognostic role of microRNA-145 in prostate cancer: A systems review and meta-analysis","volume":"3","author":"Zhang","year":"2015","journal-title":"Prostate Int."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"256","DOI":"10.1038\/sj.bjc.6605742","article-title":"The functional significance of microRNA-145 in prostate cancer","volume":"103","author":"Zaman","year":"2010","journal-title":"Br. J. Cancer"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"637","DOI":"10.1002\/pros.23156","article-title":"miR-24 regulates CDKN1B\/p27 expression in prostate cancer","volume":"76","author":"Lynch","year":"2016","journal-title":"Prostate"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1186\/s13000-020-01063-2","article-title":"High miR-324-5p expression predicts unfavorable prognosis of gastric cancer and facilitates tumor progression in tumor cells","volume":"16","author":"Zheng","year":"2021","journal-title":"Diagn. Pathol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"3220","DOI":"10.1080\/15384101.2015.1084445","article-title":"Targeting CDK6 in cancer: State of the art and new insights","volume":"14","author":"Tadesse","year":"2015","journal-title":"Cell Cycle"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1016\/j.trecan.2016.11.006","article-title":"The strange case of CDK4\/6 inhibitors: Mechanisms, resistance, and combination strategies","volume":"3","author":"Knudsen","year":"2017","journal-title":"Trends Cancer"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"123","DOI":"10.5534\/wjmh.230237","article-title":"Secondary Cancer after Androgen Deprivation Therapy in Prostate Cancer: A Nationwide Study","volume":"43","author":"Kim","year":"2025","journal-title":"World J. Men\u2019s Health"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/j.cell.2004.12.035","article-title":"Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets","volume":"120","author":"Lewis","year":"2005","journal-title":"Cell"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1139","DOI":"10.1038\/nsmb.2115","article-title":"Weak seed-pairing stability and high target-site abundance decrease the proficiency of lsy-6 and other miRNAs","volume":"18","author":"Garcia","year":"2011","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"D222","DOI":"10.1093\/nar\/gkab1079","article-title":"miRTarBase update 2022: An informative resource for experimentally validated miRNA\u2013target interactions","volume":"50","author":"Huang","year":"2022","journal-title":"Nucleic Acids Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1038\/nprot.2008.211","article-title":"Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources","volume":"4","author":"Huang","year":"2009","journal-title":"Nat. Protoc."}],"container-title":["Pharmaceuticals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8247\/19\/8\/1141\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,25]],"date-time":"2026-07-25T04:25:11Z","timestamp":1784953511000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8247\/19\/8\/1141"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,23]]},"references-count":27,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2026,8]]}},"alternative-id":["ph19081141"],"URL":"https:\/\/doi.org\/10.3390\/ph19081141","relation":{},"ISSN":["1424-8247"],"issn-type":[{"value":"1424-8247","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,23]]}}}