{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,27]],"date-time":"2025-09-27T00:07:27Z","timestamp":1758931647637,"version":"3.44.0"},"reference-count":32,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2025,9,26]],"date-time":"2025-09-26T00:00:00Z","timestamp":1758844800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"JSPS KAKENHI","award":["21K08880"],"award-info":[{"award-number":["21K08880"]}]}],"content-domain":{"domain":["www.mdpi.com"],"crossmark-restriction":true},"short-container-title":["Cancers"],"abstract":"<jats:p>Background\/Objectives: Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) is a well-established, minimally invasive method for sampling mediastinal and hilar lymph nodes. It has high sensitivity and specificity for diagnosing lymph node involvement. However, achieving a definitive cytopathological diagnosis can be challenging owing to the limited presence of atypical tumor cells. The scanning single-molecule counting (SSMC) method enables rapid measurement of target molecule expression. This study assessed the correlation between miR-200c expression measured by SSMC and lymph node metastasis confirmed by cytopathology. Methods: Following EBUS-TBNA in patients with lung cancer or suspected lung cancer, we flushed 22-gauge biopsy needles with 1 mL saline and extracted microRNA (miRNA) from the lavage fluid. The quality of SSMC results for miR-200c was evaluated and compared with that of quantitative real-time polymerase chain reaction (RT-qPCR). Results: Linear regression analysis of miR-200c between SSMC and RT-qPCR showed a statistically significant positive correlation (R2 = 0.81, p &lt; 0.0001), demonstrating the feasibility of SSMC for evaluating miRNA in lymph nodes. Based on these findings, we enrolled and analyzed 100 lymph nodes from 86 patients. High miR-200c expression detected lymph node metastasis with high sensitivity (85.7%) and specificity (83.3%) and an AUC of 0.88. No discordance was observed between the standard SSMC and rapid methods. Conclusions: The expression of miR-200c, as measured by SSMC, may serve as a biomarker for molecular nodal staging. The rapid SSMC method provides results within 30 min, significantly faster than conventional RT-qPCR, and may complement rapid on-site cytological evaluation.<\/jats:p>","DOI":"10.3390\/cancers17193133","type":"journal-article","created":{"date-parts":[[2025,9,26]],"date-time":"2025-09-26T11:28:32Z","timestamp":1758886112000},"page":"3133","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Highly Sensitive Detection of miR-200c in Metastatic Lymph Nodes Using Scanning Single-Molecule Counting"],"prefix":"10.3390","volume":"17","author":[{"given":"Yuki","family":"Sata","sequence":"first","affiliation":[{"name":"Department of General Thoracic Surgery, Graduate School of Medicine, Chiba University, Chiba 260-8670, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5585-1601","authenticated-orcid":false,"given":"Terunaga","family":"Inage","sequence":"additional","affiliation":[{"name":"Department of General Thoracic Surgery, Graduate School of Medicine, Chiba University, Chiba 260-8670, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4937-5378","authenticated-orcid":false,"given":"Takahiro","family":"Nakajima","sequence":"additional","affiliation":[{"name":"Department of General Thoracic Surgery, Dokkyo Medical University, Mibu, Shimotsuga, Tochigi 321-0293, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5885-2095","authenticated-orcid":false,"given":"Yuki","family":"Ito","sequence":"additional","affiliation":[{"name":"Department of General Thoracic Surgery, Graduate School of Medicine, Chiba University, Chiba 260-8670, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ichiro","family":"Yoshino","sequence":"additional","affiliation":[{"name":"Department of General Thoracic Surgery, Graduate School of Medicine, Chiba University, Chiba 260-8670, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8983-433X","authenticated-orcid":false,"given":"Hidemi","family":"Suzuki","sequence":"additional","affiliation":[{"name":"Department of General Thoracic Surgery, Graduate School of Medicine, Chiba University, Chiba 260-8670, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,26]]},"reference":[{"key":"ref_1","first-page":"394","article-title":"Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries","volume":"68","author":"Bray","year":"2018","journal-title":"CA Cancer J. 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