NUF2对泛癌的预后和免疫治疗效果的预测价值

doi:10.12122/j.issn.1673-4254.2025.01.17

摘要/Abstract

摘要：

目的探讨NUF2表达水平与肿瘤预后的关系及在肿瘤微环境中的作用。方法综合利用人类蛋白质图谱（HPA）、癌症基因组图谱（TCGA）、基因型-组织表达计划（GTEx）、癌细胞系百科全书（CCLE）、TIMER的数据集，采用生物信息学方法挖掘NUF2的潜在致癌作用，包括剖析NUF2在各种类型肿瘤组织中的表达水平、预后价值及免疫价值；采用RT-qPCR、Western blotting、免疫组化（IHC）等实验检测NUF2在肝癌细胞系和肝癌患者癌组织及血液中的表达情况。利用基因本体论（GO）、基因组百科全书（KEGG）和基因集富集分析（GSEA）探索NUF2及相关基因的分子机制并在肝癌中构建了NUF2的竞争性内源RNA（ceRNA）网络。结果与正常组织相比，NUF2表达在27种肿瘤中上调并与多种类型肿瘤的临床分期相关（P<0.05）。在部分肿瘤中，NUF2的表达上调与肿瘤患者的不良预后（总体生存期、疾病特异性生存期、无病生存期和无进展生存期）相关（P<0.05）。NUF2表达水平与多种肿瘤的肿瘤突变负荷、微卫星不稳定、浸润免疫细胞（B细胞、髓样树突状细胞、中性粒细胞、CD4⁺T细胞）、免疫细胞maker基因（CCR7、CD163、CD19、CD4、CD79A、CD8A、CD8B、IRF5、ITGAX和MS4A4A）以及免疫检查点基因（CTLA4、HAVCR2、LAG3、PDCD1和PDCD1LG2）呈正相关（P<0.05）。RT-qPCR等实验结果显示NUF2在肝癌细胞系、肝癌患者癌组织和血液中表达上调（P<0.05），其中肝癌患者经手术治疗之后，使得血液中NUF2表达下调（P<0.05）。GO 和 KEGG 富集分析表明，NUF2相关基因与染色体分离、微管运动以及细胞周期相关（P<0.05）。GSEA分析提示，在肝癌中NUF2高表达与甘氨酸、丝氨酸和苏氨酸代谢以及调控神经活性配体受体相互作用有关（P<0.05）。结论 NUF2在27种肿瘤中表达上调并与部分肿瘤的临床分期、不良预后有关。此外NUF2的表达与多种肿瘤的免疫细胞浸润密切相关，可能作为这些癌症免疫治疗效果的潜在预测因子。

关键词: NUF2, 泛癌分析, 免疫浸润, 预后

Abstract:

Objective To investigate the association of NUF2 expression with tumor prognosis and its regulatory role in tumor microenvironment. Methods We analyzed NUF2 expression, its prognostic value, and is immune-related functions across different cancer types using datasets from the Human Protein Atlas (HPA), TCGA, GTEx, CCLE, and TIMER. RT-qPCR, Western blotting, and immunohistochemistry were used to detect NUF2 expression in liver cancer cell lines and tissue and blood samples from patients with liver cancer. GO, KEGG, and GSEA analyses were conducted to explore the molecular mechanisms of NUF2 and its related genes, and a competitive endogenous RNA (ceRNA) network for NUF2 in liver cancer was constructed. Results NUF2 expression was upregulated in the tumor tissues of 27 cancers and was associated with clinical stages in several cancers. High NUF2 expressions were correlated with poor overall survival, disease-specific survival, progression-free survival, and disease-free survival of cancer patients. NUF2 expression levels were positively correlated with tumor mutational burden, microsatellite instability, infiltrating immune cells, immune cell marker genes and immune checkpoint genes in different cancers. RT-qPCR, Western blotting, and immunohistochemistry confirmed that NUF2 expression was upregulated in liver cancer cell lines and tumor tissues and blood samples of liver cancer patients, and was decreased significantly after operation. GO, KEGG and GSEA analyses indicated that NUF2 was involved in chromosome segregation and cell cycle and was associated with glycine, serine and threonine metabolism. Conclusion NUF2 expression is upregulated in 27 cancers and is associated with clinical stage and poor prognosis in some malignancies. NUF2 expression is closely correlated with immune cell infiltration in different cancers, suggesting its potential value for predicting immunotherapy response in these cancers.

Key words: NUF2, pan-cancer analysis, immune infiltration, prognosis

王耀彬, 陈柳燕, 罗伊凌, 申继清, 周素芳. NUF2对泛癌的预后和免疫治疗效果的预测价值[J]. 南方医科大学学报, 2025, 45(1): 137-149.

Yaobin WANG, Liuyan CHEN, Yiling LUO, Jiqing SHEN, Sufang ZHOU. Predictive value of NUF2 for prognosis and immunotherapy responses in pan-cancer[J]. Journal of Southern Medical University, 2025, 45(1): 137-149.

图/表 10

表1 qRT-PCR检测基因表达的引物序列

Tab.1 Primer sequence for qRT-PCR

Gene	Primer sequence 5'-3'
NUF2	F: AGTTAAACGCCGCACACCAG
NUF2	R: CCCTCTTGCAGCACTATCGTT
ACTIN	F: CCAACCGCGAGAAGATGACC
ACTIN	R: GAGTCCATCACGATGCCAGT

图1 分析NUF2在泛癌及不同临床分期的表达

Fig.1 Expression of NUF2 in pan-cancer and in different clinical stages. A: NUF2 mRNA expression levels in different human organs based on data from The Human Protein Atlas (www.proteinatlas.org). B: NUF2 expression in different cancers in TCGA and GTEx databases. C: NUF2 mRNA expressions in different cell lines based on CCLE database. D: Association between NUF2 expression and tumor stage. *P<0.05, **P<0.01, ***P<0.001.

图2 NUF2表达与患者总生存期的关系分析

Fig.2 Association of NUF2 expression with overall survival (OS) of cancer patients. A: Forest map of univariate Cox regression analysis of NUF2 in TCGA pan-cancer samples. B-K: Kaplan-Meier analysis of the association between NUF2 expression and OS.

图3 NUF2表达与患者无病生存期的关系分析

Fig.3 Association of NUF2 expression level with disease-free survival of cancer patients. A: Forest map of univariate Cox regression analysis of NUF2 in TCGA pan-cancer samples. B-H: Kaplan-Meier analysis of the association between NUF2 expression and DFS.

图4 NUF2表达与患者疾病特异性生存期的关系分析

Fig.4 Association of NUF2 expression with disease-specific survival (DSS) of cancer patients. A: Forest map of univariate Cox regression analysis of NUF2 in TCGA pan-cancer samples. B-K: Kaplan-Meier analysis of the association between NUF2 expression and DSS.

图5 NUF2表达与患者无进展生存期的关系分析

Fig.5 Association of NUF2 expression with progression-free survival (PFS) of cancer patients. A: Forest map of univariate Cox regression analysis of NUF2 in TCGA pan-cancer samples. B-N: Kaplan-Meier analysis of the association between NUF2 expression and PFS.

图6 NUF2表达与肿瘤突变负荷和微卫星不稳定性的关系分析

Fig.6 Association of NUF2 expression with tumor mutation burden (TMB) and microsatellite instability (MSI). A: TMB. B: MSI. *P<0.05, **P<0.01, ***P<0.001 by Spearman correlation test.

图7 NUF2与泛癌免疫细胞浸润和免疫相关基因的相关性分析

Fig.7 Correlation of NUF2 with pan-cancer immune cell infiltration and immune-related genes. A, B: Correlations of NUF2 expression with infiltration levels of various immune cells based on TIMER (A) and CIBERSORT (B) algorithms. C: Correlations of NUF2 with expressions of immune cell marker genes. D: Correlations of NUF2 with expressions of immune checkpoints-related genes. Statistical correlations were assessed using Spearman correlation test.

图8 NUF2在肝癌细胞系和患者癌组织及血液中的表达

Fig.8 NUF2 expression in HCC cell lines and tissue and blood samples from HCC patients. A, B: Levels of NUF2 in LO2 and HepG2 cells detected by RT-qPCR and Western blotting. C: Expression of NUF2 in HCC tissues detected by immunohistochemistry (n=10). D: Expression of NUF2 in HCC patients (n=29) and healthy individuals (n=15). E: Expression of NUF2 before and after surgical treatment in patients with HCC (n=14). Wilcoxon test was used to examine the significance of variations between two sets of data. *P<0.05, **P<0.01, ****P<0.0001.

图9 NUF2的分子功能及在肝癌中ceRNA调控模式预测

Fig.9 Molecular function of NUF2 and prediction of ceRNA regulatory patterns in HCC. A: Protein-protein interaction network of NUF2 based on the STRING database. B: GO and KEGG functional enrichment of NUF2-related genes. C: KEGG pathway analysis of NUF2 in GSEA in LIHC. D: miRNAs targeting NUF2 in HCC. E: Correlation analysis between hsa-miR-22-3p expression and NUF2 expression. F: Potential ceRNA regulatory axis of NUF2 in HCC. The red nodes indicate NUF2, the green nodes indicate miRNA, and the purple nodes indicate lncRNA. Statistical correlations were assessed using Spearman correlation test.

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