{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T06:01:52Z","timestamp":1789970512240,"version":"4.0.1"},"reference-count":38,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T00:00:00Z","timestamp":1789948800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003787","name":"Natural Science Foundation of Hebei Province","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100003787","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Lymph node metastasis (LNM) is a critical factor contributing to poor prognosis in esophageal squamous cell carcinoma (ESCC) patients. Consequently, elucidating its molecular mechanisms and identifying effective therapeutic targets have become a paramount challenge in clinical practice.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Plasma samples were systematically collected from healthy individuals, ESCC patients without LNM, and those with LNM. Extracellular vesicles (EVs) were isolated from plasma by size-exclusion chromatography (SEC) and ultracentrifugation (UC) methods, and EV-derived miRNA expression profiles were detected and compared by sequencing. Differentially expressed miR-1246 was selected, and a diagnostic model based on plasma EV-miR-1246 was established to predict LNM. The biological functions of EV-miR-1246 and its target gene PPP2CB, which was identified by mass spectrometry combined with online target prediction methods, in the LNM of ESCC were further investigated using\n                      <jats:italic>in vitro<\/jats:italic>\n                      cellular assays and\n                      <jats:italic>in vivo<\/jats:italic>\n                      animal models. Rescue experiments and NF-\u03baB agonist functional assays were performed to verify the downstream regulatory axis.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      MiR-1246 levels were significantly elevated in plasma extracellular vesicles from ESCC patients with lymph node metastasis. The diagnostic model established based on vesicular miR-1246 showed favorable predictive efficacy for lymph node metastasis status in ESCC.\n                      <jats:italic>In vitro<\/jats:italic>\n                      functional experiments confirmed that vesicular miR-1246 derived from ESCC cells significantly enhanced the migration, invasion, and tube formation capacities of human lymphatic endothelial cells (HLECs).\n                      <jats:italic>In vivo<\/jats:italic>\n                      mouse experiments further confirmed that vesicular miR-1246 derived from ESCC cells significantly promoted lymph node metastasis of esophageal squamous cell carcinoma in mice. Mechanistically, miR-1246 targets and represses PPP2CB expression. Rescue experiments further confirmed that the miR-1246\/PPP2CB axis activates NF-\u03baB signaling, thereby promoting lymphatic endothelial activation and lymphangiogenesis.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>This study demonstrates that EV-derived miR-1246 acts as a promising predictive biomarker for lymph-node metastasis in ESCC. Mechanistically, EV-delivered miR-1246 targets PPP2CB to activate NF-\u03baB signaling, thereby driving lymphatic endothelial activation and lymph node metastasis. These findings offer novel diagnostic biomarkers and therapeutic targets for ESCC patients with lymph node metastasis.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fimmu.2026.1881297","type":"journal-article","created":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T05:36:40Z","timestamp":1789969000000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["EV-derived miR-1246 promotes lymph node metastasis in esophageal squamous cell carcinoma through targeting the PPP2CB\/NF-\u03baB axis"],"prefix":"10.3389","volume":"17","author":[{"given":"Yaojie","family":"Wang","sequence":"first","affiliation":[{"name":"Department of Gastrocolorectal Surgery, Tongliao People\u2019s Hospital","place":["Tongliao, China"]},{"name":"Research Center, The Fourth Hospital of Hebei Medical University","place":["Shijiazhuang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuting","family":"Sun","sequence":"additional","affiliation":[{"name":"Department of Gastrocolorectal Surgery, Tongliao People\u2019s Hospital","place":["Tongliao, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuhui","family":"Fu","sequence":"additional","affiliation":[{"name":"Research Center, The Fourth Hospital of Hebei Medical University","place":["Shijiazhuang, China"]},{"name":"Key Laboratory of Tumor Gene Diagnosis, Prevention and Therapy; Clinical Oncology Research Center","place":["Shijiazhuang, Hebei, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zishuan","family":"Wei","sequence":"additional","affiliation":[{"name":"Key Laboratory of Tumor Gene Diagnosis, Prevention and Therapy; Clinical Oncology Research Center","place":["Shijiazhuang, Hebei, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lianmei","family":"Zhao","sequence":"additional","affiliation":[{"name":"Research Center, The Fourth Hospital of Hebei Medical University","place":["Shijiazhuang, China"]},{"name":"Key Laboratory of Tumor Gene Diagnosis, Prevention and Therapy; Clinical Oncology Research Center","place":["Shijiazhuang, Hebei, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guoyou","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Gastrocolorectal Surgery, Tongliao People\u2019s Hospital","place":["Tongliao, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,9,21]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.3322\/caac.21834","article-title":"Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries","volume":"74","author":"Bray","year":"2024","journal-title":"CA Cancer J Clin"},{"key":"B2","doi-asserted-by":"publisher","first-page":"176","DOI":"10.1038\/s41698-025-00929-2","article-title":"Unraveling the power of radiomics: prediction and exploration of lymph node metastasis in stage T1\/2 esophageal squamous cell carcinoma","volume":"9","author":"Zhang","year":"2025","journal-title":"NPJ Precis Oncol"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.7150\/thno.42559","article-title":"Inter- and intratumor DNA methylation heterogeneity associated with lymph node metastasis and prognosis of esophageal squamous cell carcinoma","volume":"10","author":"Teng","year":"2020","journal-title":"Theranostics"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1038\/s41571-025-01074-2","article-title":"Clinical relevance of extracellular vesicles in cancer - therapeutic and diagnostic potential","volume":"22","author":"Greening","year":"2025","journal-title":"Nat Rev Clin Oncol"},{"key":"B5","doi-asserted-by":"publisher","first-page":"18","DOI":"10.1016\/j.semcancer.2024.07.003","article-title":"Extracellular vesicles proteins for early cancer diagnosis: From omics to biomarkers","author":"De Giorgis","year":"2024","journal-title":"Semin Cancer Biol"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1002\/mco2.377","article-title":"Identification of combinatorial miRNA panels derived from extracellular vesicles as biomarkers for esophageal squamous cell carcinoma","volume":"4","author":"Wang","year":"2023","journal-title":"MedComm"},{"key":"B7","doi-asserted-by":"publisher","first-page":"1106","DOI":"10.1038\/s41416-024-02820-6","article-title":"Correction to: Serum microRNA expression profile: miR-1246 as a novel diagnostic and prognostic biomarker for oesophageal squamous cell carcinoma","volume":"131","author":"Takeshita","year":"2024","journal-title":"Br J Cancer"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1016\/j.biocel.2018.01.005","article-title":"Non-genomic mechanisms of protein phosphatase 2A (PP2A) regulation in cancer","volume":"96","author":"Kauko","year":"2018","journal-title":"Int J Biochem Cell Biol"},{"key":"B9","doi-asserted-by":"publisher","first-page":"295","DOI":"10.1186\/s13046-025-03560-y","article-title":"Targeting PP2A in cancer: an underrated option","volume":"44","author":"Sarais","year":"2025","journal-title":"J Exp Clin Cancer Res"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1038\/s41417-024-00811-6","article-title":"NF-\u03baB role on tumor proliferation, migration, invasion and immune escape","volume":"31","author":"Khan","year":"2024","journal-title":"Cancer Gene Ther"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1186\/1756-9966-30-85","article-title":"Different patterns of NF-\u03baB and Notch1 signaling contribute to tumor-induced lymphangiogenesis of esophageal squamous cell carcinoma","volume":"30","author":"Su","year":"2011","journal-title":"J Exp Clin Cancer Res"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1016\/j.redox.2020.101834","article-title":"Sustained IKK\u03b2 phosphorylation and NF-\u03baB activation by superoxide-induced peroxynitrite-mediated nitrotyrosine modification of B56\u03b33 and PP2A inactivation","volume":"41","author":"Yee","year":"2021","journal-title":"Redox Biol"},{"key":"B13","doi-asserted-by":"publisher","first-page":"10851","DOI":"10.1038\/s41598-022-14211-z","article-title":"A comparative analysis of extracellular vesicles (EVs) from human and feline plasma","volume":"12","author":"Wynne","year":"2022","journal-title":"Sci Rep"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.3390\/cells13080682","article-title":"Tumor microenvironment modulation by cancer-derived extracellular vesicles","volume":"13","author":"Ten","year":"2024","journal-title":"Cells"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.3390\/ijms22073374","article-title":"Extracellular vesicles and their role in the spatial and temporal expansion of tumor-immune interactions","volume":"22","author":"Lipinski","year":"2021","journal-title":"Int J Mol Sci"},{"key":"B16","doi-asserted-by":"publisher","first-page":"296","DOI":"10.1016\/j.semcancer.2022.06.001","article-title":"Extracellular vesicles in cancer therapy","volume":"86","author":"Wang","year":"2022","journal-title":"Semin Cancer Biol"},{"key":"B17","doi-asserted-by":"publisher","first-page":"9","DOI":"10.1016\/j.canlet.2013.02.036","article-title":"Phosphatase: PP2A structural importance, regulation and its aberrant expression in cancer","volume":"335","author":"Seshacharyulu","year":"2013","journal-title":"Cancer Lett"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1007\/s10549-017-4403-5","article-title":"Functional importance of PP2A regulatory subunit loss in breast cancer","volume":"166","author":"Watt","year":"2017","journal-title":"Breast Cancer Res Treat"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.3791\/61178","article-title":"Blocking lymph flow by suturing afferent lymphatic vessels in mice","volume":"23","author":"Lin","year":"2020","journal-title":"J Vis Exp"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbc.2021.101380","article-title":"Histone deacetylase 5 deacetylates the phosphatase PP2A for positively regulating NF-\u03baB signaling","volume":"297","author":"Xu","year":"2021","journal-title":"J Biol Chem"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1126\/science.aau6977","article-title":"The biology, function, and biomedical applications of exosomes","volume":"367","author":"Kalluri","year":"2020","journal-title":"Science"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1016\/bs.ctm.2024.06.010","article-title":"The role of extracellular vesicles in cancer","volume":"94","author":"Perez Hurtado","year":"2024","journal-title":"Curr Top Membr"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1016\/j.canlet.2025.217735","article-title":"Extracellular vesicles: Hermes between cancers and lymph nodes","volume":"623","author":"Cao","year":"2025","journal-title":"Cancer Lett"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.3389\/fonc.2021.721785","article-title":"Role of extracellular vesicles on cancer lymphangiogenesis and lymph node metastasis","volume":"11","author":"Wang","year":"2021","journal-title":"Front Oncol"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.3390\/ijms27031509","article-title":"Clinical applications of extracellular vesicles: Promises and pitfalls","volume":"27","author":"Primorac","year":"2026","journal-title":"Int J Mol Sci"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1038\/s43018-021-00272-y","article-title":"Melanoma-derived small extracellular vesicles induce lymphangiogenesis and metastasis through an NGFR-dependent mechanism","volume":"2","author":"Garc\u00eda-Silva","year":"2021","journal-title":"Nat Cancer"},{"key":"B27","doi-asserted-by":"publisher","first-page":"95","DOI":"10.2147\/IJN.S391173","article-title":"Exosomal miR-10527-5p inhibits migration, invasion, lymphangiogenesis and lymphatic metastasis by affecting Wnt\/\u03b2-catenin signaling via Rab10 in esophageal squamous cell carcinoma","volume":"18","author":"Xiao","year":"2023","journal-title":"Int J Nanomedicine"},{"key":"B28","doi-asserted-by":"publisher","first-page":"4309","DOI":"10.3390\/cancers16234039","article-title":"The lymphatic vascular system in extracellular vesicle-mediated tumor progression","volume":"16","author":"Lodha","year":"2024","journal-title":"Cancers (Basel)"},{"key":"B29","doi-asserted-by":"publisher","first-page":"13","DOI":"10.1186\/s11658-022-00410-z","article-title":"Exosomal circ_0026611 contributes to lymphangiogenesis by reducing PROX1 acetylation and ubiquitination in human lymphatic endothelial cells (HLECs)","volume":"28","author":"Yao","year":"2023","journal-title":"Cell Mol Biol Lett"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1177\/15353702211021022","article-title":"Regulation of lymphangiogenesis by extracellular vesicles in cancer metastasis","volume":"246","author":"Wang","year":"2021","journal-title":"Exp Biol Med"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1002\/jev2.12341","article-title":"Tumour extracellular vesicles induce neutrophil extracellular traps to promote lymph node metastasis","volume":"12","author":"Su","year":"2023","journal-title":"J Extracell Vesicles"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1002\/1878-0261.13029","article-title":"Circulating exosomal miR-363-5p inhibits lymph node metastasis by downregulating PDGFB and serves as a potential noninvasive biomarker for breast cancer","volume":"15","author":"Wang","year":"2021","journal-title":"Mol Oncol"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1038\/nm.2753","article-title":"Melanoma exosomes educate bone marrow progenitor cells toward a pro-metastatic phenotype through MET","volume":"18","author":"Peinado","year":"2012","journal-title":"Nat Med"},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-25-0663","article-title":"Extracellular vesicle-packaged miR-99a reprograms fibroblasts to create an inflammatory niche that drives colorectal cancer metastasis","volume":"86","author":"Zhou","year":"2026","journal-title":"Cancer Res"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.adv6937","article-title":"AKR1B10 dictates c-Myc stability to suppress colorectal cancer metastasis via PP2A nitration","volume":"11","author":"Wu","year":"2025","journal-title":"Sci Adv"},{"key":"B36","doi-asserted-by":"publisher","first-page":"56","DOI":"10.1016\/j.jphs.2022.07.001","article-title":"PP2A regulates metastasis and vasculogenic mimicry formation via PI3K\/AKT\/ZEB1 axis in non-small cell lung cancers","volume":"150","author":"Zhang","year":"2022","journal-title":"J Pharmacol Sci"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1080\/15476286.2019.1585738","article-title":"The origin of exosomal miR-1246 in human cancer cells","volume":"16","author":"Xu","year":"2019","journal-title":"RNA Biol"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1021\/acs.bioconjchem.1c00068","article-title":"New lipophilic fluorescent dyes for labeling extracellular vesicles: Characterization and monitoring of cellular uptake","volume":"32","author":"Shimomura","year":"2021","journal-title":"Bioconjug Chem"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2026.1881297\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T05:36:41Z","timestamp":1789969001000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2026.1881297\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,21]]},"references-count":38,"alternative-id":["10.3389\/fimmu.2026.1881297"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2026.1881297","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,21]]},"article-number":"1881297"}}