Lv Tianyun, Zhu Ziyan, Xin Yunchao, Liu Yaping, Chang Zhicheng, Yin Xiaoxia, Shang Xiaoling
Otorhinolaryngology, Head and Neck Surgery, Hebei North University, Zhangjiakou, Hebei, China.
Otorhinolaryngology, Head and Neck Surgery,The First Affiliated Hospital of Hebei North University, Zhangjiakou, Hebei, China.
Genet Test Mol Biomarkers. 2025 Jun;29(6):152-165. doi: 10.1089/gtmb.2025.0046. Epub 2025 Jun 2.
Herein, we addressed the clinical challenge of high lymph node metastasis rates despite the lack of reliable diagnostic biomarkers in papillary thyroid carcinoma (PTC) by employing bioinformatics approaches to identify key biomarkers, aiming to provide new strategies for clinical diagnosis and treatment. Through bioinformatics analysis, plasminogen activator urokinase () was identified as a key biomarker for lymph node metastasis in PTC. Immunohistochemistry (IHC) was performed to validate expression in tumor and adjacent normal tissues and its correlation with clinicopathological features. cellular expression was further confirmed by immunocytochemistry (ICC), Western blotting, and quantitative real-time PCR (qRT-PCR). Cell Counting Kit-8 and Transwell assays were used to assess its role in PTC tumor cell proliferation, migration, and invasion. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed on -related genes; immune cell infiltration in PTC was evaluated using the TIMER database and CIBERSORT algorithm. Bioinformatics analysis showed that expression was significantly elevated in the PTC lymph node metastasis group [area under the receiver operating characteristic curve, 75.3%]. IHC results demonstrated significantly elevated expression in tumor tissues. Clinicopathological correlation analysis indicated that was associated with lymph node metastasis, particularly lateral cervical lymph node involvement. ICC, qRT-PCR, and Western blotting confirmed that was highly expressed in PTC tumor cells. After transient knockdown of , proliferation, migration, and invasion of PTC tumor cells were significantly reduced. GO and KEGG enrichment analyses showed that -related genes were primarily involved in signal transduction, inflammatory response, and P53, PI3K-Akt, and Mitogen-activated protein kinase (MAPK) signaling pathways. Immune cell infiltration was significantly higher in PTC tissues than in adjacent normal tissues; expression positively correlated with B and CD8 T cell infiltration and Programmed cell death protein 1 (PD-1) and Cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) expression. enhances PTC cell proliferation, migration, and invasion while promoting immune escape through the Th1/Th2 imbalance and PD-1/CTLA-4 upregulation, serving as a potential biomarker for lymph node metastasis in PTC.
在此，我们通过采用生物信息学方法来识别关键生物标志物，应对了甲状腺乳头状癌（PTC）中尽管缺乏可靠诊断生物标志物但淋巴结转移率较高的临床挑战，旨在为临床诊断和治疗提供新策略。通过生物信息学分析，尿激酶型纤溶酶原激活剂（uPA）被确定为PTC中淋巴结转移的关键生物标志物。进行免疫组织化学（IHC）以验证uPA在肿瘤组织和相邻正常组织中的表达及其与临床病理特征的相关性。通过免疫细胞化学（ICC）、蛋白质免疫印迹法和定量实时聚合酶链反应（qRT-PCR）进一步证实了uPA的细胞表达。使用细胞计数试剂盒-8和Transwell实验来评估其在PTC肿瘤细胞增殖、迁移和侵袭中的作用。对与uPA相关的基因进行基因本体论和京都基因与基因组百科全书富集分析；使用TIMER数据库和CIBERSORT算法评估PTC中的免疫细胞浸润情况。生物信息学分析表明，uPA表达在PTC淋巴结转移组中显著升高[受试者操作特征曲线下面积，75.3%]。IHC结果显示肿瘤组织中uPA表达显著升高。临床病理相关性分析表明，uPA与淋巴结转移相关，尤其是颈外侧淋巴结受累。ICC、qRT-PCR和蛋白质免疫印迹法证实uPA在PTC肿瘤细胞中高表达。短暂敲低uPA后，PTC肿瘤细胞的增殖、迁移和侵袭显著降低。基因本体论和京都基因与基因组百科全书富集分析表明，与uPA相关的基因主要参与信号转导、炎症反应以及P53、PI3K-Akt和丝裂原活化蛋白激酶（MAPK）信号通路。PTC组织中的免疫细胞浸润显著高于相邻正常组织；uPA表达与B细胞和CD8 + T细胞浸润以及程序性细胞死亡蛋白1（PD-1）和细胞毒性T淋巴细胞相关蛋白4（CTLA-4）表达呈正相关。uPA增强PTC细胞增殖、迁移和侵袭，同时通过Th1/Th2失衡和PD-1/CTLA-4上调促进免疫逃逸，作为PTC中淋巴结转移的潜在生物标志物。
