{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T22:59:44Z","timestamp":1790204384884,"version":"4.1.0"},"reference-count":22,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T00:00:00Z","timestamp":1790208000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Genet."],"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Post-radiotherapy chronic pharyngitis affects 60%\u201380% of laryngeal cancer survivors and is characterized by sustained elevation of pro-inflammatory cytokines and dysregulation of key apoptotic and oncogenic signaling pathways, including NF-\u03baB\/PI3K-AKT\/EGFR and TP53\/CASP3 axes. Modified Bixuesan (MBS), a classical Traditional Chinese Medicine formula with nearly 180 years of clinical use in pharyngolaryngeal disorders, has not been systematically characterized for its molecular mechanism of action in this context.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      We conducted an integrative study combining network pharmacology, multi-omics bioinformatics, and\n                      <jats:italic>in vitro<\/jats:italic>\n                      cell line validation. Active MBS compounds were screened from TCMSP and BATMAN-TCM databases using ADME thresholds. Single-cell transcriptomic data from laryngeal squamous cell carcinoma (LSCC; GSE127519) and bulk microarray data from radiation-induced pharyngitis (GSE103412) were analyzed to identify hub target genes. Molecular docking was performed using AutoDock Vina against crystal structures from RCSB PDB.\n                      <jats:italic>In vitro<\/jats:italic>\n                      RT-qPCR validation was conducted in Hep-2 and TU212 laryngeal cancer cell lines treated with MBS extract (0.5\u00a0mg\/mL, 48\u00a0h), evaluating mRNA expression of four hub genes: TP53, EGFR, NF-\u03baB1, and CASP3.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Network pharmacology identified 47 active MBS compounds and 89 putative therapeutic targets, with TP53, AKT1, EGFR, NF-\u03baB1, CASP3, MMP9, and VEGFA as principal hub genes. Molecular docking revealed strong predicted binding of quercetin to TP53 (\u22129.4\u00a0kcal\/mol) and kaempferol to EGFR (\u22128.7\u00a0kcal\/mol).\n                      <jats:italic>In vitro<\/jats:italic>\n                      RT-qPCR in MBS-treated Hep-2 cells demonstrated significant upregulation of TP53 (1.72 \u00b1 0.19-fold, p = 0.024) and CASP3 (1.89 \u00b1 0.25-fold, p = 0.011), and significant downregulation of EGFR (0.46 \u00b1 0.08-fold, p = 0.003) and NF-\u03baB1 (0.43 \u00b1 0.11-fold, p = 0.004). In TU212 cells, concordant and more pronounced changes were observed: TP53 (2.51 \u00b1 0.31-fold, p = 0.001), CASP3 (2.24 \u00b1 0.27-fold, p = 0.002), EGFR (0.38 \u00b1 0.07-fold, p &amp;lt; 0.001), and NF-\u03baB1 (0.36 \u00b1 0.09-fold, p = 0.001).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      This integrative study provides\n                      <jats:italic>in vitro<\/jats:italic>\n                      gene expression evidence suggesting that MBS may modulate inflammatory and apoptotic signaling associated with post-radiotherapy laryngeal mucosal injury, through predicted regulation of NF-\u03baB\/PI3K-AKT\/EGFR pathway components and upregulation of TP53\/CASP3 mRNA expression. These findings are exploratory; network pharmacology and docking analyses are hypothesis-generating only, and\n                      <jats:italic>in vitro<\/jats:italic>\n                      results require prospective mechanistic and clinical validation.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fgene.2026.1917203","type":"journal-article","created":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T22:10:17Z","timestamp":1790201417000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Modified Bixuesan modulates inflammatory and apoptotic gene expression in post-radiotherapy chronic pharyngitis of laryngeal cancer: an integrative network pharmacology and in vitro validation study"],"prefix":"10.3389","volume":"17","author":[{"given":"Dongqing","family":"Lu","sequence":"first","affiliation":[{"name":"Department of Traditional Chinese Medicine, Beicai Community Health Service Center","place":["Shanghai, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuxu","family":"Guo","sequence":"additional","affiliation":[{"name":"Department of Traditional Chinese Medicine, Shanghai Tianchen Rehabilitation Hospital","place":["Shanghai, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jun","family":"Gao","sequence":"additional","affiliation":[{"name":"Department of General Practice, Beicai Community Health Service Center","place":["Shanghai, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanming","family":"Cai","sequence":"additional","affiliation":[{"name":"Department of General Practice, Beicai Community Health Service Center","place":["Shanghai, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaofei","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Comprehensive Treatment, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine","place":["Shanghai, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lulu","family":"Jiao","sequence":"additional","affiliation":[{"name":"Department of Traditional Chinese Medicine, Beicai Community Health Service Center","place":["Shanghai, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fei","family":"He","sequence":"additional","affiliation":[{"name":"Department of Traditional Chinese Medicine, Beicai Community Health Service Center","place":["Shanghai, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,9,24]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"229","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. 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