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. 2024 May 29:14:1399484.
doi: 10.3389/fonc.2024.1399484. eCollection 2024.

IRX5's influence on macrophage polarization and outcome in papillary thyroid cancer

Lu Qin #  1 , Cheng Chen #  2 , Zhengwei Gui  3 , Yun Jiang  4
Affiliations

IRX5's influence on macrophage polarization and outcome in papillary thyroid cancer

Lu Qin et al. Front Oncol. .

Abstract

Background: With a rise in recent years, thyroid cancer (TC) is the most prevalent hormonal cancer worldwide. It is essential to investigate the inherent variability at the molecular level and the immune environment within tumors of various thyroid cancer subtypes in order to identify potential targets for therapy and provide precise treatment for patients with thyroid adenocarcinoma.

Methods: First, we analyzed the expression of IRX5 in pan-cancer and papillary thyroid carcinoma by bioinformatics methods and collected paired samples from our center for validation. Subsequently, we analyzed the significance of IRX5 on the prognosis and diagnosis of PTC. Next, we explored the possible mechanisms by which IRX5 affects the prognosis of thyroid cancer patients by GO/KEGG enrichment analysis, and further investigated the effect of IRX5 on immune infiltration of thyroid cancer. Ultimately, by conducting experiments on cells and animals, we were able to show how IRX5 impacts the aggressive characteristics of thyroid cancer cells and its influence on macrophages within the immune system of thyroid cancer.

Results: In 11 malignant tumors, including PTC, IRX5 is overexpressed and associated with a poor prognosis. IRX5 may affect the prognosis of PTC through embryonic organ development, ossification, mesenchyme development, etc. Increased IRX5 expression decreases the presence of cytotoxic and Th17 cells in papillary thyroid cancer. IRX5 was highly expressed in different PTC cell lines, such as K-1 and TPC-1. Silencing IRX5 effectively halted the growth and movement of PTC cells while also decreasing M2 polarization and enhancing M1 polarization in tumor-associated macrophages.

Conclusion: IRX5 could impact the outlook of individuals with PTC by stimulating the shift of macrophages to M2 in the immune surroundings of thyroid cancer growths, suggesting a potential new focus for treating thyroid cancer, particularly through immunotherapy.

Keywords: IRX5 gene; macrophage polarization; proliferation; thyroid cancer; tumor immune microenvironment.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
The expression difference of IRX5 in cancer tissue and normal tissue. (A) Expression of IRX5 in unpaired thyroid cancer samples in TCGA-THTC database. (B) Expression of IRX5 in paired thyroid cancer samples in TCGA-THTC database. (C) Expression of IRX5 in pan-cancer and adjacent normal tissues in TCGA and GTEx databases. Data were shown as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001.
Figure 2
Figure 2
Verification of IRX5 overexpression in thyroid cancer in our center specimen. (A) Typical cancer and paracancer IRX5 immunohistochemical images. (B) Quantitative IRX5 immunohistochemical analysis of 10 paired specimens. *p < 0.05.
Figure 3
Figure 3
Expression of IRX5 and prognosis of thyroid cancer patients. (A) Diagnostic ROC curve of IRX5. (B) Time-dependent ROC curve of IRX5. (C) OS of thyroid cancer patients based on IRX5 expression level.
Figure 4
Figure 4
Analysis of IRX5-related enrichment pathways. (A) Heatmap of TOP25 genes positively associated with IRX5 co-expression. (B) Heatmap of TOP25 genes negatively associated with IRX5 co-expression. (C) Wayne plot of IRX5 co-expressed genes taking intersection with PTC survival related genes. (D) GO/KEGG analysis of IRX5 co-expressed genes. (E) GO/KEGG analysis of intersecting genes. (F) Protein interactions network of intersecting genes.
Figure 5
Figure 5
Associated between IRX5 with immune cell infiltration. (A) Correlation between the expression level of IRX5 and various immune cell infiltration. (B) Correlation between IRX5 expression and Eosinophils. (C) Correlation between IRX5 expression and Mast cells. (D) Correlation between IRX5 expression and NK cells. (E) correlation between IRX5 expression and Th17 cells. ns, not statistically.
Figure 6
Figure 6
Expression and knockdown of IRX5 in various cell lines and CCK8 cell proliferation experiment. (A) IRX5 expression in Nthy-ori3–1, BCPAP, FTC-133, K-1 and TPC-1 cell lines. (B) IRX5 knockdown efficiency of two siRNA in K-1 cell lines. (C) IRX5 knockdown of two siRNA in TPC-1 cell lines Efficiency. (D, E) Cell proliferation in two siRNA knockout groups and control groups in K-1 and TPC-1 cell lines. **p < 0.01, ***p < 0.001.
Figure 7
Figure 7
Cellular and animal experiments to validate the effect of IRX5 on PTC cells. (A) Clone formation of control group and two siRNA knockout groups in K-1 and TPC-1 cell lines. (B) Scratch test images of control group and two siRNA knockout groups in K-1 and TPC-1 cell lines. (C) Transwell images of control group and two siRNA knockout groups in K-1 and TPC-1 cell lines. (D) Quantitative analysis of clone formation experiment. (E) Quantitative analysis of scratch experiment. All assays were independently repeated at least three times. (F) Quantitative analysis of transwell experiment. (G) Growth of transplanted tumors in nude mice injected with K-1 cells and si-IRX5 K-1 cells. (H) Weight of grafted tumors. Data are presented as the mean ± SD. **p < 0.01, ***p < 0.001.
Figure 8
Figure 8
Effect of IRX5 knockdown on thyroid cancer tumor-associated macrophages. (A) Schematic diagram of the co-culture model of thyroid cancer cells and macrophages. (B) Cytokine content (ELISA) of culture fluid in co-culture chambers. (C) Macrophage polarization marker expression. (D) Immunofluorescence graph of macrophage morphology (si-IRX5). (E) Immunofluorescence graph of macrophage morphology (si-Ctrl). (F) Detection of macrophage polarization in mouse tumors by flow cytometry. * p < 0.05, **p < 0.01, ***p < 0.001. ns, not statistically.
Figure 9
Figure 9
Immune cell infiltration in TCGA-THCA. (A) TIMER scores related to IRX5 expression in TCGA-THCA (the cohort was divided into two groups by the 50% median IRX5 expression. G1:IRX5 high expression group, G2:IRX5 low expression group) (B) TCIA database analysis of the M1 to M2 ratio of TAM in various malignancies. ns, not statistically. *p < 0.05, ***p < 0.001, ****p < 0.0001.
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References

    1. Burman KD, Wartofsky L. CLINICAL PRACTICE. thyroid nodules. N Engl J Med. (2015) 373(24):2347–56. doi: 10.1056/NEJMcp1415786 - DOI - PubMed
    1. Pellegriti G, Frasca F, Regalbuto C, Squatrito S, Vigneri R. Worldwide increasing incidence of thyroid cancer: update on epidemiology and risk factors. J Cancer Epidemiol. (2013) 2013:965212. doi: 10.1155/2013/965212 - DOI - PMC - PubMed
    1. Song M, Sun W, Liu Q, Wang Z, Zhang H. Global scientific trends on thyroid disease in early 21st century: a bibliometric and visualized analysis. Front Endocrinol (Lausanne). (2023) 14:1306232. doi: 10.3389/fendo.2023.1306232 - DOI - PMC - PubMed
    1. Gomez-Skarmeta JL, Modolell J. Iroquois genes: genomic organization and function in vertebrate neural development. Curr Opin Genet Dev. (2002) 12(4):403–8. doi: 10.1016/S0959-437X(02)00317-9 - DOI - PubMed
    1. Zhu L, Dai L, Yang N, Liu M, Ma S, Li C, et al. . Transcription factorIRX5 promotes hepatocellular carcinoma proliferation and inhibits apoptosis by regulating the p53 signalling pathway. Cell Biochem Funct. (2020) 38(5):621–9. doi: 10.1002/cbf.3517 - DOI - PMC - PubMed

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