Graduate School, Beijing University of Chinese Medicine.
Oncology Department, Wangjing Hospital, China Academy of Chinese Medical Sciences.
J Vis Exp. 2024 Sep 20(211). doi: 10.3791/67421.
This study aimed to probe the potential common pathogenic mechanisms linking primary Sjogren's syndrome (pSS) and lung adenocarcinoma (LUAD) through bioinformatics analysis and experimental verification. The relevant genes associated with pSS and LUAD were retrieved from the Gene Expression Omnibus (GEO) database and Genecard database. Subsequently, differentially expressed genes (DEGs) associated with pSS and LUAD were screened as pSS-LUAD-DEGs. Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment analyses were performed to elucidate the significant biological functions of pSS-LUAD-DEGs. Core targets were identified by constructing the protein-protein interaction (PPI) network, further assessing hub gene diagnostic accuracy through Receiver Operating Characteristic (ROC) curve analyses. In this study, NOD/Ltj mice served as pSS animal models and were stimulated with particulate matter 2.5 (PM2.5) to generate an inflammatory reaction. Quantitative real-time polymerase chain reaction (qPCR), enzyme-linked immunosorbent assay (ELISA), and western blotting were employed for relevant molecular biology experiment verification. The results revealed through KEGG and GO enrichment analyses indicate that inflammation plays a critical role in linking pSS and LUAD. IL6, CCNA2, JAK2, IL1B, ASPM, CCNB2, NUSAP1, and CEP55 were determined as key targets of pSS-LUAD. BALB/c mice and NOD/Ltj mice exhibited enhanced expression of inflammatory cytokines IL-6 and IL-1β in lung tissues following 21 days of stimulation with PM2.5, activating the JAK2/STAT3 signaling pathway and up-regulating the expression of tumor-associated genes CCNA2, CCNB2, and CEP55, with NOD/Ltj mice exhibiting more pronounced changes than BALB/c mice. This protocol demonstrates that carcinogenesis induced by the pulmonary inflammatory microenvironment may be a key reason for the high incidence of LUAD in pSS patients. Additionally, blocking-related mechanisms may help prevent the occurrence of LUAD in pSS patients.
本研究旨在通过生物信息学分析和实验验证，探究原发性干燥综合征（pSS）和肺腺癌（LUAD）之间潜在的共同致病机制。从基因表达综合数据库（GEO）和基因卡片数据库（Genecard）中检索与 pSS 和 LUAD 相关的基因。随后，筛选与 pSS 和 LUAD 相关的差异表达基因（DEGs）作为 pSS-LUAD-DEGs。进行京都基因与基因组百科全书（KEGG）和基因本体论（GO）富集分析，以阐明 pSS-LUAD-DEGs 的重要生物学功能。通过构建蛋白质-蛋白质相互作用（PPI）网络确定核心靶点，进一步通过接受者操作特征（ROC）曲线分析评估关键基因的诊断准确性。在这项研究中，NOD/Ltj 小鼠被用作 pSS 动物模型，并受到细颗粒物 2.5（PM2.5）的刺激以产生炎症反应。采用定量实时聚合酶链反应（qPCR）、酶联免疫吸附测定（ELISA）和蛋白质印迹法进行相关分子生物学实验验证。通过 KEGG 和 GO 富集分析揭示的结果表明，炎症在连接 pSS 和 LUAD 方面起着关键作用。IL6、CCNA2、JAK2、IL1B、ASPM、CCNB2、NUSAP1 和 CEP55 被确定为 pSS-LUAD 的关键靶点。BALB/c 小鼠和 NOD/Ltj 小鼠在 PM2.5 刺激 21 天后，肺组织中炎症细胞因子 IL-6 和 IL-1β 的表达增强，激活 JAK2/STAT3 信号通路并上调肿瘤相关基因 CCNA2、CCNB2 和 CEP55 的表达，NOD/Ltj 小鼠的变化比 BALB/c 小鼠更为明显。该方案表明，肺部炎症微环境诱导的癌变可能是 pSS 患者 LUAD 高发病率的一个关键原因。此外，阻断相关机制可能有助于预防 pSS 患者发生 LUAD。
