目的·系统鉴定RNA结合蛋白人类抗原R（human antigen R，HuR）在非小细胞肺癌（non-small cell lung cancer，NSCLC）中的下游关键靶基因，并阐明其通过调控该靶点影响肿瘤进展的分子机制。方法·基于癌症基因组图谱（The Cancer Genome Atlas，TCGA）数据库、基因型-组织表达项目（Genotype-Tissue Expression，GTEx）数据库及临床蛋白质组学肿瘤分析联盟（Clinical Proteomic Tumor Analysis Consortium，CPTAC）数据库，分析HuR在NSCLC及癌旁组织中的表达差异；通过免疫组织化学法检测HuR蛋白水平，并利用GSE19188数据集进行验证。基于TCGA数据，分析HuR表达与临床病理参数间关系，进行生存分析，并通过单因素与多因素Cox回归评估预后因素，构建列线图模型预测生存率。利用ESTIMATE包及CIBERSORT算法分析HuR表达与免疫微环境的相关性。采用Transwell实验检测HuR敲低后A549细胞迁移与侵袭能力变化。通过RNA测序筛选差异表达基因（differentially expressed genes，DEGs），结合蛋白质相互作用（protein-protein interaction，PPI）网络鉴定枢纽基因，并利用基因本体论（Gene Ontology，GO）与京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes，KEGG）进行富集分析。采用RNA免疫沉淀（RNA immunoprecipitation，RIP）验证HuR与整合素β1（integrin β 1，ITGB1） mRNA的直接结合，并通过实时定量PCR（RT-qPCR）和Western blotting分别检测HuR敲低后ITGB1在mRNA和蛋白水平的表达变化。结果·HuR在肺癌组织中显著高表达，与不良预后独立相关（P=0.045），且其表达与免疫抑制性微环境特征呈负相关。功能实验表明敲低HuR可显著抑制肺癌细胞的迁移（P<0.001）与侵袭（P=0.002）能力。生物信息学分析将ITGB1锁定为HuR下游的核心枢纽基因，富集分析显示其参与细胞外基质受体相互作用等通路。RIP实验证实HuR可直接结合ITGB1 mRNA；进一步的RT-qPCR与Western blotting结果表明，敲低HuR导致ITGB1在mRNA和蛋白水平均显著下调，提示HuR主要通过维持ITGB1 mRNA稳定性发挥转录后调控作用。结论·HuR通过直接结合并稳定ITGB1 mRNA，激活下游信号通路，促进NSCLC进展。HuR-ITGB1调控轴的发现不仅为深入理解肺癌的发病机制提供了新视角，也为预后判断和靶向治疗提供了潜在新靶点。

Objective ·To systematically identify the key downstream target genes of the RNA-binding protein human antigen R (HuR) in non-small cell lung cancer (NSCLC) and to elucidate the molecular mechanisms through which HuR influences tumor progression by regulating this target. Methods ·Based on the The Cancer Genome Atlas (TCGA), Genotype-Tissue Expression (GTEx), and Clinical Proteomic Tumor Analysis Consortium (CPTAC) databases, the differential expression of HuR in NSCLC and adjacent tissues was analyzed. HuR protein levels were detected via immunohistochemistry and validated by using the GSE19188 dataset. Using TCGA data, the relationship between HuR expression and clinicopathological parameters was analyzed; survival analysis was performed, and univariate and multivariate Cox regression analyses were conducted to assess prognostic factors, followed by the construction of a nomogram model to predict survival rates. The ESTIMATE package and CIBERSORT algorithm were used to analyze the correlation between HuR expression and the tumor immune microenvironment. A Transwell assay was employed to detect changes in the migration and invasion capabilities of A549 cells after HuR knockdown. RNA-seq was used to screen for differentially expressed genes (DEGs), and hub genes were identified by combining protein-protein interaction (PPI) network analysis. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed for functional enrichment. Mechanistically, RNA immunoprecipitation (RIP) was used to validate the direct binding between HuR and integrin β 1 (ITGB1) mRNA. Furthermore, real-time quantitative PCR (RT-qPCR) and Western blotting were performed to detect the expression changes of ITGB1 at the mRNA and protein levels, respectively, after HuR knockdown. Results ·HuR was significantly overexpressed in lung cancer tissues, independently associated with poor prognosis (P=0.045), and negatively correlated with characteristics of an immunosuppressive microenvironment. Functional experiments demonstrated that HuR knockdown significantly inhibited the migration (P<0.001) and invasion (P=0.002) capabilities of lung cancer cells. Bioinformatic analysis identified ITGB1 as the core hub gene downstream of HuR, and enrichment analysis revealed its significant involvement in pathways such as the extracellular matrix (ECM)-receptor interaction. RIP assays confirmed that HuR directly binds to ITGB1 mRNA. Further RT-qPCR and Western blotting results indicated that HuR knockdown led to significant downregulation of ITGB1 at both the mRNA (P=0.001) and protein levels, suggesting that HuR primarily exerts its post-transcriptional regulatory role by maintaining ITGB1 mRNA stability. Conclusion ·HuR promotes NSCLC progression by directly binding to and stabilizing ITGB1 mRNA, thereby activating downstream signaling pathways. The discovery of the HuR-ITGB1 regulatory axis not only provides a novel perspective for understanding the pathogenesis of lung cancer but also offers a potential target for prognosis assessment and targeted therapy.