我们在The Cancer Genome Atlas（TCGA）的Genomic Data Commons（GDC）门户网站中分析了头颈部鳞状细胞癌（HNSCC）患者的RNA测序（RNA-seq）和临床数据，研究了无粘系相关基因（ARGs）在HNSCC中的预后价值，并开发了新的靶向药物。使用生物信息学方法筛选出差异表达的ARGs；随后构建了含有三个ARGs（CDKN2A，BIRC5和PLAU）的预后模型。我们的结果表明，基于模型的风险评分是一个良好的预后指标，并通过TISCH数据库验证了三个ARGs在HNSCC预后中的潜力，模型的准确性在Gene Expression Omnibus数据库的两个独立队列中得到了验证。还进行了免疫相关分析和半最大抑制浓度分析，揭示了风险组之间不同的时间风景，并预测免疫和化疗反应。随后使用L1000FWD数据库预测了HNSCC的潜在小分子药物。最后，采用体外实验验证了数据库的发现。HNSCC内相对ARG mRNA表达水平与模型结果保持一致。BIRC5沉默抑制了WSU-HN6和CAL-27细胞中的无粘抵抗。分子对接、实时PCR、细胞计数试剂盒-8（CCK-8）、脉冲克隆和流式细胞术分析显示，数据库预测的小分子药物可能针对预后模型中的ARGs，抑制HNSCC细胞的存活率，并在体外促进无粘。因此，我们构建了一个用于HNSCC患者的新的ARG模型，可以预测预后和免疫活性，并确定潜在的HNSCC小分子药物，为临床上针对HNSCC中的无粘铺平了道路。© 2023 Qiu等。

We analyzed RNA-sequencing (RNA-seq) and clinical data from head and neck squamous cell carcinoma (HNSCC) patients in The Cancer Genome Atlas (TCGA) Genomic Data Commons (GDC) portal to investigate the prognostic value of anoikis-related genes (ARGs) in HNSCC and develop new targeted drugs. Differentially expressed ARGs were screened using bioinformatics methods; subsequently, a prognostic model including three ARGs (CDKN2A, BIRC5, and PLAU) was constructed. Our results showed that the model-based risk score was a good prognostic indicator, and the potential of the three ARGs in HNSCC prognosis was validated by the TISCH database, the model's accuracy was validated in two independent cohorts of the Gene Expression Omnibus database. Immune correlation analysis and half-maximal inhibitory concentration were also performed to reveal the different landscapes of TIME between risk groups and to predict immuno- and chemo-therapeutic responses. Potential small-molecule drugs for HNSCC were subsequently predicted using the L1000FWD database. Finally, in vitro experiments were used to verify the database findings. The relative ARG mRNA expression levels in HNSCC and surrounding normal tissues remained consistent with the model results. BIRC5 knockdown inhibited anoikis resistance in WSU-HN6 and CAL-27 cells. Molecular docking, real-time PCR, cell counting kit-8 (CCK-8), plate clone, and flow cytometry analyses showed that small-molecule drugs predicted by the database may target the ARGs in the prognostic model, inhibit HNSCC cells survival rate, and promote anoikis in vitro. Therefore, we constructed a new ARG model for HNSCC patients that can predict prognosis and immune activity and identify a potential small-molecule drug for HNSCC, paving the way for clinically targeting anoikis in HNSCC.© 2023 Qiu et al.