Shi Ling, Qiu Feng, Shi Chao, Zhang Guohua, Yu Feng
Department of Oncology, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, 7889 Changdong Avenue, Gaoxin District, Nanchang, 330006, Jiangxi Province, China.
Department of Oncology, First People's Hospital of Jiujiang City, Jiujiang, 332000, Jiangxi Province, China.
Discov Oncol. 2025 Aug 12;16(1):1541. doi: 10.1007/s12672-025-02529-9.
Non-small cell lung cancer (NSCLC) is one of the most prevalent and deadliest cancers worldwide, accounting for a significant global health burden. Targeted therapies such as osimertinib, a third-generation EGFR inhibitor, have transformed the treatment landscape for EGFR-mutant NSCLC by offering improved progression-free survival. However, the inevitable development of resistance remains a formidable challenge, necessitating deeper insights into its molecular underpinnings. In this study, we employed an integrative bioinformatics approach to analyze multi-cohort transcriptomic datasets, uncovering 126 resistance-associated genes, revealing 50 significant osimertinib resistance-related genes, and identifying eight key hub genes (KRT14, KRT16, KRT17, KRT5, KRT6A, KRT6B, TP63, and TRIM29) that contribute to immune evasion and tumor microenvironment remodeling. Integrated qPCR and Western blot analyses validated the significant upregulation of KRT14, KRT16, KRT6A, and TRIM29 in osimertinib-resistant cell lines (PC9 OR and HCC827 OR) at both transcriptional and translational levels, with KRT14 exhibiting the most pronounced upregulation. Functional assays demonstrated that KRT14 knockdown restored osimertinib sensitivity, suppressed proliferation, and impaired migration in resistant cells. Functional enrichment analyses revealed critical pathways, including p53 signaling and metabolic reprogramming, underlying resistance mechanisms. Batch effect analysis highlighted a marked reduction in effector immune cells, such as activated CD8 + T cells, alongside an increase in immunosuppressive populations, emphasizing the role of immune evasion in osimertinib resistance.We constructed a robust diagnostic model, nomoScore, based on the hub genes, achieving excellent predictive accuracy (AUC > 0.9) in training and validation datasets. These findings offer novel insights into resistance mechanisms and propose actionable strategies for integrating targeted and immunotherapies to improve outcomes for NSCLC patients. Future experimental and clinical studies are essential to validate and translate these findings into therapeutic advances.
非小细胞肺癌（NSCLC）是全球最常见且最致命的癌症之一，给全球健康带来了沉重负担。靶向治疗，如第三代EGFR抑制剂奥希替尼，通过提高无进展生存期改变了EGFR突变型NSCLC的治疗格局。然而，耐药性的不可避免发展仍然是一个巨大挑战，需要更深入了解其分子基础。在本研究中，我们采用综合生物信息学方法分析多队列转录组数据集，发现了126个与耐药相关的基因，揭示了50个与奥希替尼耐药显著相关的基因，并确定了八个关键枢纽基因（KRT14、KRT16、KRT17、KRT5、KRT6A、KRT6B、TP63和TRIM29），这些基因有助于免疫逃逸和肿瘤微环境重塑。综合定量PCR和蛋白质印迹分析在转录和翻译水平验证了奥希替尼耐药细胞系（PC9 OR和HCC827 OR）中KRT14、KRT16、KRT6A和TRIM29的显著上调，其中KRT14上调最为明显。功能测定表明，敲低KRT14可恢复奥希替尼敏感性，抑制耐药细胞增殖并损害其迁移。功能富集分析揭示了包括p53信号传导和代谢重编程在内的关键途径，这些途径是耐药机制的基础。批次效应分析突出显示效应免疫细胞，如活化的CD8 + T细胞显著减少，同时免疫抑制群体增加，强调了免疫逃逸在奥希替尼耐药中的作用。我们基于枢纽基因构建了一个强大的诊断模型nomoScore，在训练和验证数据集中实现了出色的预测准确性（AUC > 0.9）。这些发现为耐药机制提供了新见解，并提出了整合靶向治疗和免疫治疗以改善NSCLC患者预后的可行策略。未来的实验和临床研究对于验证这些发现并将其转化为治疗进展至关重要。
