人乳头瘤病毒（HPV）阴性头颈鳞状细胞癌（HNSCC）的分子分类仍然存在疑问。在肿瘤和正常组织之间检测到差异表达的基因，GSEA 显示它们与细胞周期途径相关。本研究旨在根据细胞周期相关基因对 HPV 阴性 HNSCC 进行分类。已建立的基因模式与肿瘤进展、临床预后和药物治疗功效相关。使用从癌症基因组图谱 (TCGA)、临床蛋白质组肿瘤分析联盟 (CPTAC) 和基因表达综合 (GEO) 数据库获得的 HNSCC 患者样本数据进行生物学分析。本研究中包含的所有样本都包含生存信息。 740 个样本的 RNA 测序数据用于分析。先前表征的细胞周期相关基因被纳入无监督共识聚类中。鉴定出 HPV 阴性 HNSCC 的两种亚型（C1、C2）。 C1亚型表现出低细胞周期活性、“热”肿瘤微环境（TME）、较早的N分期、较低的病理分级、较好的预后以及对免疫治疗和靶向治疗的较高反应率。 C2 亚型与较高的细胞周期活性、“冷”TME、较晚的 N 分期、较高的病理分级、较差的预后和较低的治疗反应率相关。根据最近模板预测方法，建立分类规则并进行验证。我们的工作从细胞周期的角度探讨了HPV阴性HNSCC的分子机制，可能为个体化抗癌治疗提供新的思路。版权所有：©2023 Gu et al.这是一篇根据知识共享署名许可条款分发的开放获取文章，允许在任何媒体上不受限制地使用、分发和复制，前提是注明原始作者和来源。

The molecular classification of human papillomavirus (HPV)-negative head and neck squamous cell carcinomas (HNSCCs) remains questionable. Differentially expressed genes were detected between tumor and normal tissues and GSEA showed they are associated with cell cycle pathways. This study aimed to classify HPV-negative HNSCCs based on cell cycle-related genes. The established gene pattern was correlated with tumor progression, clinical prognosis, and drug treatment efficacy. Biological analysis was performed using HNSCC patient sample data obtained from the Cancer Genome Atlas (TCGA), Clinical Proteomic Tumor Analysis Consortium (CPTAC), and Gene Expression Omnibus (GEO) databases. All samples included in this study contained survival information. RNA sequencing data from 740 samples were used for the analysis. Previously characterized cell cycle-related genes were included for unsupervised consensus clustering. Two subtypes of HPV-negative HNSCCs (C1, C2) were identified. Subtype C1 displayed low cell cycle activity, 'hot' tumor microenvironment (TME), earlier N stage, lower pathological grade, better prognosis, and higher response rate to the immunotherapy and targeted therapy. Subtype C2 was associated with higher cell cycle activity, 'cold' TME, later N stage, higher pathological grade, worse prognosis, and lower response rate to the treatment. According to the nearest template prediction method, classification rules were established and verified. Our work explored the molecular mechanism of HPV-negative HNSCCs in the view of cell cycle and might provide new sights for personalized anti-cancer treatment.Copyright: © 2023 Gu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.