Objective:After selecting NCF2 based on bioinformatics, clinical experiments were conducted to verify the expression of NCF2 in chronic rhinosinusitis with nasal polyps to study its correlation. Methods:The differentially expressed genes（DEGs） between CRSwNP and non-CRS patients were explored using the CRS-related dataset from the gene expression omnibus GEO database. The weighted gene co-expression network（WGCNA） was used for cluster analysis. The expression and cell distribution of NCF2 in the tissues were determined by single gene enrichment analysis（GSEA）, immune inflammatory infiltration analysis, and principal component（PCA） analysis. The expression degree of NCF2 in the tissues of the subjects was determined by immunohistochemistry, and the percentage of EOS in the peripheral blood of the subjects was detected and the correlation was analyzed. EOS in the tissues of the subjects were counted under a microscope and compared. Results:①The Venn diagram was obtained by crossing the module with the highest correlation between DEGs and WGCNA to determine the core gene NCF2. ②GSEA analysis showed that NCF2 was significantly related to the immunological processes such as allogeneic rejection and asthma. ③The area under the ROC curve was 1, indicating that NCF2 had diagnostic value for CRSwNP. ④NCF2 was highly expressed in nasal polyps, mainly distributed in monocytes and eosinophils. ⑤HE staining showed that the number of EOS in ECRSwNP tissues and the percentage of eosinophils in peripheral blood were higher than those in nonECRSwNP and control groups. ⑥The immunohistochemistry results showed that NCF2 was significantly expressed in the nasal polyps of ECRSwNP patients, which was higher than that in the nasal mucosa of nonECRSwNP group and control group. ⑦The expression of NCF2 in tissues was positively correlated with EOS count in ECRSwNP group and EOS expression in peripheral blood. Conclusion:The expression of NCF2 is increased in eosinophilic chronic rhinosinusitis with nasal polyps, and it is significantly correlated with the expression of eosinophils in peripheral blood and tissues, suggesting that NCF2 may be used as a basis for the intrinsic classification of ECRSwNP and a reference index for clinical diagnosis and treatment.
目的：基于生物信息学选定NCF2后通过临床实验验证NCF2在慢性鼻窦炎伴鼻息肉（chronic sinusitis with nasal polyps, CRSwNP）中表达从而研究其相关性。 方法：使用基因表达综合数据库GEO中CRS相关数据集挖掘CRSwNP与非CRS患者之间差异表达基因（DEGs），加权基因共表达网络（WGCNA）进行聚类分析，WGCNA相关性最高模块与DEGs相交确定核心基因NCF2，通过单基因富集分析（GSEA）、免疫炎性浸润分析、主成分（PCA）分析确定NCF2在组织中表达情况及细胞分布情况，运用免疫组织化学实验测定NCF2在研究对象组织中表达程度，同时检测研究对象外周血中EOS百分比并进行相关性分析，镜下计数研究对象组织中嗜酸粒细胞（EOS）并进行比较分析。 结果：①DEGs结果与WGCNA相关性最高模块相交得到韦恩图，确定核心基因NCF2；②GSEA分析显示NCF2与异体排斥反应、哮喘等免疫反应过程显著相关；③ROC曲线下面积为1，显示NCF2对CRSwNP具有诊断价值；④免疫炎性浸润及PCA分析显示NCF2在鼻息肉组织中高表达，主要分布在单核细胞及EOS；⑤苏木精-伊红染色结果显示ECRSwNP组织中EOS计数及外周血中EOS的百分比均高于非ECRSwNP（nonECRSwNP）组与对照组；⑥免疫组织化学结果显示NCF2在EOS型CRSwNP（eosinophilic CRSwNP, ECRSwNP）患者鼻息肉组织中表达显著，高于nonECRSwNP组和对照组患者鼻腔黏膜中表达水平；⑦NCF2在组织中的表达与ECRSwNP组中的EOS计数及外周血中EOS表达均呈正相关。 结论：NCF2在CRSwNP组织中表达增高，与外周血及组织中EOS表达相关性显著，提示NCF2或可作为CRSwNP的内在分型依据及临床诊疗参考。.
Keywords: NCF2; bioinformatics analysis; chronic rhinosinusitis with nasal polyps; eosinophils.
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