Predictive value of NUF2 for prognosis and immunotherapy responses in pan-cancer

doi:10.12122/j.issn.1673-4254.2025.01.17

Abstract

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

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.

Figures/Tables 10

Tab.1 Primer sequence for qRT-PCR

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

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.

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.

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.

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.

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.

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.

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.

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.

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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