MUC5B regulates alterations in the immune microenvironment in nasopharyngeal carcinoma via the Wnt/β-catenin signaling pathway

Abstract

Objective: To screen potential differentially expressed genes related to immune function in nasopharyngeal carcinoma through an online database, and to verify their mechanism of action, so as to provide a reference for the diagnosis and treatment of nasopharyngeal carcinoma in the future.

Methods: Differentially expressed genes were analyzed from the GSE227541 dataset, and functional enrichment analysis was conducted. With mucin 5B, oligomeric mucus/gel-forming as the focus, the correlation between its expression and immune indexes was analyzed by using the TIMER database. The expressions of mucin 5B, oligomeric mucus/gel-forming in clinical NPC tissues and adjacent tissue samples were detected Furthermore, mucin 5B, oligomeric mucus/gel-forming abnormal expression vectors were constructed and transfected into human NPC cell CNE-2Z to detect alterations in cell activity, ferroptosis and the immune microenvironment. In addition, the role of the Wnt/β-catenin signaling pathway in nasopharyngeal carcinoma was observed, and the influence of mucin 5B, oligomeric mucus/gel-forming on this pathway was discussed.

Results: A total of 42 differentially expressed genes were found in the GSE227541 dataset, among which mucin 5B, oligomeric mucus/gel-forming was obviously at the core of the entire network. In nasopharyngeal carcinoma tissues, the research team observed the upregulated expression of mucin 5B, oligomeric mucus/gel-forming (P < 0.05). In vitro, increased expression of elevated mucin 5B, oligomeric mucus/gel-forming activates nasopharyngeal carcinoma cell activity and immune escape and inhibits ferroptosis. In terms of pathways, upregulating mucin 5B, oligomeric mucus/gel-forming expression could activate the Wnt/β-catenin pathway.

Conclusions: Mucin 5B, oligomeric mucus/gel-forming regulates the immune escape of nasopharyngeal carcinoma cells and participates in tumor progression by mediating the Wnt/β-catenin signaling pathway.

Keywords: Bioinformatics; Immunization; Mucin 5B; Nasopharyngeal carcinoma; Oligomeric mucus/gel-forming; Wnt signaling pathway; β-catenin.

Fig. 1

Online database screening for DEGs in NPC. A Expression of DEGs in the GSE227541 dataset (volcano plot). B PPI network of DEGs. C GO analysis of DEGs. (D) KEGG analysis of DEGs. Nasopharyngeal carcinoma (NPC), differentially expressed genes (DEGs), gene ontology (GO), kyoto encyclopedia of genes and genomes (KEGG), protein–protein interaction (PPI), Mucin 5B, oligomeric mucus/gel-forming (MUC5B)

Fig. 2

Relationship between MUC5B and the immune microenvironment. A MUC5B expression in various tumors. B Relationship between MUC5B and HAAD, LUAD, and HNSC immune infiltrating cells. C Impact of MUC5B on the prognosis of HNSC patients. D SCNA analysis of the effect of MUC5B on immune infiltrating cells. E Expression of MUC5B in NPC patients. ***P < 0.001. Somatic copy-number alterations (SCNA), Mucin 5B, oligomeric mucus/gel-forming (MUC5B)

Fig. 3

Effect of MUC5B on NPC cells. A Effect of interfering with MUC5B expression on ferroptosis in NPC cells. B Effect of interfering with MUC5B expression on the growth ability of NPC cells. C Effects of interfering with MUC5B expression on the proliferative capacity of NPC cells. D Effect of interfering with MUC5B expression on the invasion ability of NPC cells. E Effect of interfering with MUC5B expression on the migration ability of NPC cells. F Effect of interfering with MUC5B expression on immune infiltration of NPC cells. * and *** indicate P < 0.05 and P < 0.001, respectively, compared with the blank group; # and ### indicate P < 0.05 and P < 0.001, respectively, compared with the overexpression group. Mucin 5B, oligomeric mucus/gel-forming (MUC5B), Nasopharyngeal carcinoma (NPC), Glutathione peroxidase 4 (GPX4), Recombinant solute carrier family 7, member 11 (SLC7A11), Recombinant acyl coenzyme a synthetase long chain family, member 4 (ACSL4), 5-Ethynyl-2'-deoxyuridine (EdU), Programmed death-1 (PD-1), Programmed cell death 1 ligand 1 (PD-L1)

Fig. 4

Impact of MUC5B on NPC live tumors. A Effect of MUC5B on tumor growth in vivo (volume and weight). B Effect of MUC5B on tumor histopathological damage (HE staining). C Effect of MUC5B on CD8 + cells in NPC loaded mice. D Effect of MUC5B on immune infiltration of tumor tissues. * and *** indicate P < 0.05 and P < 0.001, respectively, compared with the blank group; # and ### indicate P < 0.05 and P < 0.001, respectively, compared with the overexpression group. Mucin 5B, oligomeric mucus/gel-forming (MUC5B), Nasopharyngeal carcinoma (NPC), Programmed death-1 (PD-1), Programmed cell death 1 ligand 1 (PD-L1), pathway inhibition (PI), combined intervention (CI)

Fig. 5

MUC5B regulates immune escape in NPC via the Wnt/β-catenin signaling pathway. A Effect of MUC5B on the expression of the Wnt/β-catenin pathway. B MUC5B regulates the growth of NPC cells through the Wnt/β-catenin pathway. C MUC5B regulates the proliferation of NPC cells through the Wnt/β-catenin pathway. D MUC5B regulates NPC cell invasion through the Wnt/β-catenin pathway. E MUC5B regulates NPC cell migration through the Wnt/β-catenin pathway. F MUC5B regulates ferroptosis of NPC cells through the Wnt/β-catenin pathway. G MUC5B regulates NPC tumor growth in vivo through the Wnt/β-catenin pathway. H MUC5B regulates pathological injury of NPC living tumor tissues through the Wnt/β-catenin pathway. * and *** indicate P < 0.05 and P < 0.001, respectively, compared with the blank group; # and ### indicate P < 0.05 and P < 0.001, respectively, compared with the PI group. Mucin 5B, oligomeric mucus/gel-forming (MUC5B), Nasopharyngeal carcinoma (NPC), Glutathione peroxidase 4 (GPX4), Recombinant solute carrier family 7, member 11 (SLC7A11), Recombinant acyl coenzyme a synthetase long chain family, member 4 (ACSL4), 5-Ethynyl-2ʹ-deoxyuridine (EdU), Programmed death-1 (PD-1), Programmed cell death 1 ligand 1 (PD-L1)