A comprehensive prognostic and immune infiltration analysis of UBA1 in pan-cancer: A computational analysis and in vitro experiments

Abstract

Ubiquitin like modifier activating enzyme 1 (UBA1) plays an important role in immune regulation and cellular function. However, the functional mechanism and role of UBA1 in pan-cancer have not been fully elucidated and its value in haematological tumours (diffuse large B cell lymphoma (DLBC/DLBCL) and acute myeloid leukaemia (AML/LAML)) has not been explored. We conducted a comprehensive analysis of the functional mechanism and role of UBA1 in pan-cancer using multiple databases, including differential expression analysis, clinical pathological staging analysis, prognosis analysis and immune analysis. Then, we confirmed the function of UBA1 in haematological tumours through cell experiments. The results showed that the expression of UBA1 was significantly increased in most cancers and the differential expression of UBA1 was mainly concentrated in digestive tumours, haematological tumours and brain tumours. Moreover, the high expression of UBA1 had poor prognosis in most tumours, which may be related to its involvement in various cancer-related pathways such as cell cycle, as well as its methylation level, protein phosphorylation level, immune cell infiltration and immune therapy response. Cell experiments have confirmed that UBA1 can significantly regulate the cycle progression and apoptosis of DLBCL cells and AML cells. Therefore, UBA1 may be a potential therapeutic target for haematological tumours. In summary, our study not only comprehensively analysed the functional mechanisms and clinical value of UBA1 in pan-cancer, but also validated for the first time the regulatory role of UBA1 in haematological tumours.

Keywords: UBA1; Haematological tumours; Pan‐cancer; cancer immunity; prognosis.

FIGURE 1

The workflow of this study.

FIGURE 2

Differential expression of UBA1 in pan‐cancer. The GEPIA2.0 results showed that UBA1 mRNA in cancer was highly expressed in BRCA, COAD, DLBC, ESCA, GBM, LAML, LGG, LIHC, OV, PAAD, READ, STAD and THYM and significantly downregulated in KIRC (FC >1.4 or <0.7 and p < 0.01).

FIGURE 3

Pan‐cancer analysis of the prognostic value of UBA1.The Kaplan Meier survival curve showed that upregulation of UBA1 expression was significantly associated with poor OS in LAML, BRCA, LGG, LIHC and LUAD (A–E).

FIGURE 4

UBA1 coexpressed gene/protein interaction network and enrichment analysis. Analysing the top 100 co‐expressed genes of UBA1 in pan‐cancer on GEPIA2.0, the top 5 genes (CDK16, ELK1, KDM5C, RBM10 and TBC1D25) were highly correlated with UBA1 in most cancer types (A). The functional enrichment of GO exhibits activities related to histone methylation, negative regulation of mRNA metabolism, negative regulation of protein modification, DNA template transcription, protein ubiquitination, and cell cycle (B). Then, further research was conducted on the potential impact of UBA1 on protein interaction networks, and 20 genes related to UBA1 were extracted from the GeneMANIA database for GO and KEGG enrichment analysis (C). GO enrichment analysis showed that UBA1 is involved in some pathways, including ubiquitin like modifier activating enzyme activity, serine family amino acid biosynthesis process, DNA damage response, and microtubule cytoskeletal tissue. In addition, KEGG pathway analysis showed that UBA1 was involved in the ubiquitin mediated protein hydrolysis pathway (D).

FIGURE 5

UBA1 and immune infiltration. UBA1 is positively and significantly correlated with matrix score, ImmuneScore and microenvironment score in many cancers. In most tumours, UBA1 expression also shows a significant positive correlation with immune cell infiltration. UBA1 showed a significant positive correlation with immune score, immune microenvironment score, Neutrophil, MacrophageM2 and Macrophage in GBM and LGG, a significant positive correlation with MacrophageM1 in GBM and a positive correlation with MacrophageM1 in LGG. There is a significant positive correlation with T cell CD4 + Th2 in GBM and a significant negative correlation with T cell CD4 + Th2 in LGG. UBA1 is significantly positively correlated with immune cell infiltration in CESC, CHOL, ESCA, LUAD, MESO, OV and SKCM.

FIGURE 6

UBA1 and immune checkpoint. In TGCT, UBA1 was significantly positively correlated with 8 immune checkpoint genes.

FIGURE 7

Cycle experiments of DLBCL cells. The results of cell proliferation showed that compared with the OCI‐LY1 + NC group cells, the OCI‐LY1 + UBA1‐Overexpression group showed a significant increase in G1 phase cells decreased, S phase cells showed no significant changes and G2 phase cells increased (A, B). Therefore, the high expression of UBA1 significantly promotes the progression of DLBCL cell cycle.

FIGURE 8

Apoptosis experiments of DLBCL cells. The results of cell apoptosis showed that compared with the OCI‐LY1 + NC group cells, the apoptosis rate of the OCI‐LY1 + UBA1‐Overexpression group cells were significantly reduced (A, B). Therefore, the high expression of UBA1 significantly inhibited apoptosis of DLBCL cells.

FIGURE 9

Cycle experiments of AML cells. The results of cell proliferation showed that compared with the HL‐60 + NC‐siRNA group cells, the HL‐60 + UBA1‐siRNA1 group showed a significant increase in G1 phase cells increase, S phase cells remain unchanged and G2 phase cells decrease (A, B). Therefore, the low expression of UBA1 significantly inhibited the progression of AML cell cycle.

FIGURE 10

Apoptosis experiments of AML cells. The results of cell apoptosis showed that compared with the HL‐60 + NC‐siRNA group cells, the apoptosis rate of the HL‐60 + UBA1‐siRNA1 group cells was significantly increased (A, B). Therefore, the low expression of UBA1 significantly promotes apoptosis of AML cells.