Abstract
Background
Morbidity and mortality rates of Digestive System Cancers (DSC) continue to pose human lives and health. Nuclear factor erythroid 2-like protein 3 (NFE2L3) is aberrantly expressed in DSC. This study aimed to explore the clinical value and underlying mechanisms of NFE2L3 as a novel biomarker in DSC.
Methods
We utilized data from databases and clinical gastric cancer specimens to validate the aberrant expression level of NFE2L3 and further assessed the clinical value of NFE2L3. To investigate the potential molecular mechanism of NFE2L3, we analyzed the correlation of NFE2L3 with immune molecular mechanisms, constructed PPI network, performed GO analysis and KEGG analysis, and finally explored the biological function of NFE2L3 in gastric cancer cells.
Results
NFE2L3 expression is up-regulated in DSC and has both prognostic and diagnostic value. NFE2L3 correlates with various immune mechanisms, PPI network suggests proteins interacting with NFE2L3, GSEA analysis suggests potential molecular mechanisms for NFE2L3 to play a role in cancer promotion, and in vitro cellular experiments also confirmed the effect of NFE2L3 on the biological function of gastric cancer cells.
Conclusion
Our study confirms the aberrant expression and molecular mechanisms of NFE2L3 in DSC, indicating that NFE2L3 could serve as a novel biomarker for diagnosis and prognosis of DSC.
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Introduction
Globally, tumors continue to pose a threat to human life and health. In 2021, statistics from the American Cancer Society showed that there were 338,090 new cases and 169,280 deaths were estimated for digestive tract tumors in the United States (Siegel et al. 2021). DSC mainly include cholangiocarcinoma (CHOL), colon adenocarcinoma (COAD), esophageal carcinoma (ESCA), liver hepatocellular carcinoma (LIHC), pancreatic adenocarcinoma (PAAD), rectum adenocarcinoma (READ), and stomach adenocarcinoma (STAD), continue to be major types of cancer threatening human health and survival. DSC are affected by several risk factors, including lifestyle habits, tobacco use, alcoholism, physical activity, and gender (** the progression of DSC. Relevant studies have indicated that cancers harbor high TMB means the potential to generate many neoantigens, making tumors more immunogenic for ICIs (Bravaccini et al. 2021; Chan et al. 2019). Expect MSI, the TMB also demonstrated the significance of the DSC. A higher TMB is normally related to an adverse outcome in advanced tumors.
According to our results, we revealed that NFE2L3 was closely correlated with immunity. It is expected to become novel immunotherapeutic target for DSC in future. Further studies should continue to investigate the mechanisms of NFE2L3 in immunotherapy.
Our findings indicated a significant positive correlation between NFE2L3 and immune or molecular subtypes. Cellular and immune subtypes are the main factors responsible for tumor heterogeneity. Treatment elicits varied responses from tumors, owing to the genetic diversity within the tumor. The combination of immune and molecular subtypes for precisely targeted tumor therapy will also greatly enhance the current status of immunotherapeutic efficacy. Therefore, we believe that NFE2L3 plays a vital role in DSC immunotherapy. Therapeutic cancer vaccines remain a valid immunotherapy option and hold significant implications for the clinical advancement of cancer patients (Pe'er et al. 2021; Baharom et al. 2022). Over the past few years, there has been a significant amount of research conducted on vaccines and therapeutics specifically designed for the innate and adaptive immune systems in the field of cancer treatment (Peng et al. 2022; Ratnam et al. 2019). Thus, the important functions of immune cells in tumor-specific immunity against immune cells can be understood. Our findings showed that compared with non-responders, patients who generated antibody responses to therapy exhibited prolonged DFS. We have reason to believe that NFE2L3 to be a potential target for immunotherapy in future.
A multitude of studies have indicated that patients with immune cell infiltration tumors exhibit a more favorable response to ICI than patients with non-immunological tumors. Recent research has revealed that NFE2L3 plays a pivotal role in cancer development (Wang et al. 2018; Zhang et al. 2019), but the precise mechanisms have yet to be elucidated. The findings of this research indicated that DSCs experienced immune cell infiltration. The immune cell infiltration encompasses a wide range of cell types, such as Tregs, T cells, dendritic cells, NK cells, T follicular helper cells, macrophages, mast cells, and monocytes. In STAD, we observed a strong correlation between NFE2L3 expression and immune cells abundance. The research revealing the correlation of immune cell infiltration with NFE2L3 may open unique avenues for precise cancer therapeutics against cancers.
Co-expression analysis can be employed as a means to identify disease-associated genes and gene functions in tumors. Chemokines combine with the cell surface chemokine receptors to perform biological functions, such as the chemotaxis, leukocyte migration, and inflammatory activities (Charo and Ransohoff 2006; Arimont et al. 2017). The causative role of chemokines is variable, and they play an important role in many diseases, such as cancer, viral infections, inflammatory, and autoimmune diseases. Researchers have found that chemokines affect anti-tumorigenic activity by regulating tumor angiogenesis and infiltration of immune cells (Lacalle et al. 2017). The results of our study showed a strong correlation between NFE2L3 and immune-related genes or chemokine receptors. In COAD, LIHC, and STAD, there was a close correlation between NFE2L3 expression and the chemokine receptors, such as C-X3-C motif chemokine receptor 1 (CX3CR1), plexin B2 (PLXNB2), atypical chemokine receptor 1 (ACKR1), and formyl peptide receptor 2 (FPR2). Anja Schmall found that the tumor-associated crosstalk between macrophages and cancer cells via the CCR2 and CX3CR1 signaling pathways directed the lung cancer growth and metastasis (Schmall et al. 2015). Circ_0013958 plays an oncogenic role in ovarian cancer by regulating the miR-637/PLXNB2 axis (Liang et al. 2021). Immune-related genes, such as PD-1, PD-L1, and CTLA4, have been successfully used in tumor immunotherapy and have been shown to have a significant effect on B cell lymphomas, HCC, and non-small cell lung cancer (Xu-Monette and Zhou 2018; Kamada et al. 2019), (Thommen et al. 2018). This study revealed novel immune-related genes that are co-expressed with NFE2L3 in DSC, including TIGIT, IDO1, ADORA2A, and CD70, which can be used as immune therapy, checkpoints, and biomarkers (Zhai et al. 2018; Chauvin and Zarour 2020). Therefore, combining immune-related genes or chemokine receptors with NFE2L3 may enhance the efficacy of DSC diagnosis and immunotherapy.
We conducted GO enrichment and KEGG analysis on the NFE2L3 binding proteins, we found that the Hedgehog signaling pathway is the primary target of protein catabolic processes and cell differentiation. Hedgehog signaling pathway activated tumors by driving EMT, and the inhibitor of Hedgehog signaling pathway exhibited remarkable clinical outcomes in various types of cancer Zhang et al. (2009). Nevertheless, the investigation into the differential regulation of target genes by NFE2L3 remains to be investigated.
NFE2L3 is involved in the regulation of various biological and cellular processes, such as cell cycle, cell differentiation, or inflammatory processes. We performed GSEA and found that NFE2L3 was significantly associated with epigenetics, methylation modification, cell cycle regulation, calcium signaling pathway and steroid hormone biosynthesis. Consequently, the distinct manifestation of NFE2L3 assumes a crucial function in DSC through various biological functions and signaling pathways, ultimately impacting the process of tumorigenesis. Nevertheless, a comprehensive understanding of the findings would require further experimental validation.
To validate the bioinformatics findings, in vivo experiments conducted on gastric cancer cells demonstrated the role of NFE2L3 as an oncogene, facilitating the proliferation and movement of gastric cancer cells. NFE2L3 enhances tumor cell migration ability by affecting the EMT through Wnt/β-catenin signaling pathway (Ren et al. 2020; Zeng and Ju 2018). These findings and our hypothesis are validated experimental results. More in vitro research is needed to gain a better understanding of how things work in future.
The outcomes of these bioinformatics analyses lay the groundwork for future comprehensive investigations into the mechanisms underlying tumorigenesis and evolution. NFE2L3 was found to be aberrantly expressed in DSC, and significantly correlated with the diagnosis and prognosis of DSC patients. In addition, in vivo results suggest that NFE2L3 may act as an oncogene roles in gastric cancer through promoting the gastric cancer cells growth and migration.
Conclusions
In conclusion, we identified a specific role of NFE2L3 in DSC. The results of our study indicated that NFE2L3 was differentially expressed in cancers and closely related to clinical features. Therefore, it has the potential prognostic and diagnostic values for DSC. In addition, NFE2L3 was closely related to immune filtration, immune subtype, and molecular subtype and co-expressed with numerous genes and chemokines. These findings suggest that NFE2L3 has the potential to emerge as a novel therapeutic target for DSC. Additional experimental studies are required to verify these findings.
Data availability
All relevant data are within the manuscript and its Additional files.
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This work was supported by the National Natural Science Foundation of China (grant numbers 82070594).
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Conception and design: Fan Li; Provision of study materials or patients: Fan Li, Zhili Wen; Collection and assembly of data: Fan Li; Data analysis: Fan Li; Manuscript writing: Fan li; Revised: Zhili Wen; Funding acquisition, Zhili Wen.
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Li, F., Wen, Z. Identification roles of NFE2L3 in digestive system cancers. J Cancer Res Clin Oncol 150, 150 (2024). https://doi.org/10.1007/s00432-024-05656-y
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DOI: https://doi.org/10.1007/s00432-024-05656-y