Wakisaka Naohiro, Moriyama-Kita Makiko, Kondo Satoru, Kobayashi Eiji, Ueno Takayoshi, Nakanishi Yosuke, Endo Kazuhira, Sugimoto Hisashi, Yoshizaki Tomokazu
Department of Otorhinolaryngology, NHO Kanazawa Medical Center, Kanazawa, Ishikawa, Japan.
Division of Otorhinolaryngology and Head and Neck Surgery, Graduate School of Medical Science, Kanazawa University, Kanazawa, Ishikawa, Japan.
PLoS One. 2025 Jan 16;20(1):e0316102. doi: 10.1371/journal.pone.0316102. eCollection 2025.
Cancer immune responses are generated in secondary lymphoid organs, such as the lymph nodes and tonsils. In the current study, transcriptional profiles of peritumoral tonsillar tissues (PTTs) from oropharyngeal cancers (OPCs) were assessed and compared with those of inflammatory tonsils and regional lymph nodes (rLNs).
RNA samples of PTTs and rLNs from 13 OPCs, and 4 inflammatory tonsils were subjected to microarray analysis, and differentially expressed genes (DEGs) identified from 730 nCounter Panel immune-related genes. Gene Set enrichment Analysis (GSEA) was used for DEG profiling of PTTs and rLNs between lymph node metastasis-negative and metastasis-positive cases. The top 20 genes, as ranked by GSEA metric scores, were extracted and subjected to principal component analysis (PCA). The correlation of each patient's PCA score with lymph node status was assessed by Receiver Operating Characteristics (ROC) analysis.
Comparing DEG analyses of PTTs with those of inflammatory tonsils and rLNs revealed 144 and 45 upregulated genes, respectively. ClueGO, a widely used Cytoscape plug-in, revealed activated pathways in PTTs, including lymphocyte proliferation (followed by T cell activation involved in the immune response) and positive regulation of leukocyte migration (followed by antimicrobial humoral immune response mediated by antimicrobial peptides) as the most significantly enriched immune system process functions in the gene ontology when comparing inflammatory tonsils and rLNs. The area under the ROC curves of PTTs and rLNs were 0.806 and 0.389, and were significant by DeLong's test (p = 0.025).
PTTs exhibit unique immunological features distinguishing them from inflammatory tonsils and rLNs. Gene expression analysis of PTTs is useful for investigating the mechanism of OPC lymphatic spread, even compared with analysis of rLNs.
癌症免疫反应在二级淋巴器官中产生,如淋巴结和扁桃体。在本研究中,评估了口咽癌(OPC)瘤旁扁桃体组织(PTT)的转录谱,并与炎性扁桃体和区域淋巴结(rLN)的转录谱进行比较。
对13例OPC的PTT和rLN以及4例炎性扁桃体的RNA样本进行微阵列分析,从730个nCounter Panel免疫相关基因中鉴定出差异表达基因(DEG)。基因集富集分析(GSEA)用于淋巴结转移阴性和转移阳性病例之间PTT和rLN的DEG分析。提取按GSEA指标评分排名的前20个基因,并进行主成分分析(PCA)。通过接受者操作特征(ROC)分析评估每位患者的PCA评分与淋巴结状态的相关性。
将PTT的DEG分析与炎性扁桃体和rLN的DEG分析进行比较,分别发现144个和45个上调基因。广泛使用的Cytoscape插件ClueGO揭示了PTT中激活的通路,包括淋巴细胞增殖(其次是参与免疫反应的T细胞激活)和白细胞迁移的正调控(其次是由抗菌肽介导的抗微生物体液免疫反应),在比较炎性扁桃体和rLN时,这是基因本体中最显著富集的免疫系统过程功能。PTT和rLN的ROC曲线下面积分别为0.806和0.389,经DeLong检验具有显著性(p = 0.025)。
PTT表现出独特的免疫学特征,使其有别于炎性扁桃体和rLN。PTT的基因表达分析对于研究OPC淋巴扩散机制很有用,甚至与rLN分析相比也是如此。