<i>ISG15</i> promotes M5-induced hacat cell proliferation through Wnt signaling in psoriasis

Xianqi Sun; Yuzhen Li; Huiwen Yu; Jiaying Lin; Chen Wang; Quanlin Liu; Bingxue Bai

doi:10.1515/fzm-2024-0022

ISG15 promotes M5-induced hacat cell proliferation through Wnt signaling in psoriasis

doi: 10.1515/fzm-2024-0022

1.
Department of Dermetology, the Second Affiliated Hospital of Harbin Medical University, Harbin 150001, China
2.
Heilongjiang University of Chinese Medicine, Ministry of Education, Harbin 150040, China

Funds:

the Natural Science Foundation of Heilongjiang Province PL2024H107

More Information

Corresponding author: Bingxue Bai, E-mail: bxbddd@163.com

摘要

Abstract: Objective Psoriasis is a common chronic, recurrent, immune-mediated inflammatory skin disease, which tends to occur in cold areas. Its pathogenesis is currently unclear. This study aims to screen differentially expressed genes in the psoriasis dataset, identify the central genes, detect the expression of central genes in psoriasis lesions of patients in the cold regions and then conduct further research. Methods Differential genes associated with psoriasis in the GEO database were analyzed, and functional enrichment analysis and protein-protein interaction network analysis. The expression results of the identified genes were validated in psoriasis cell models. The ISG15 gene, which showed the most significant difference in expression, was further studied. The expression level of ISG15 protein in psoriasis was examined. Then, we knocked out ISG15 in psoriasis cell models and detected keratinocyte proliferation by MTT, Real-Time PCR and Western Blot. Western Blot showed the expression of β-catenin after ISG15 gene knockout. Results We detected the protein expression of ISG15 in the cold area of Northeast China, and found that the expression of ISG15 increased in patients with psoriasis, and the proliferation of keratinocytes and the expression of β-catenin decreased in psoriasis cell model after ISG15 was knocked down. ISG15 regulates keratinocyte proliferation through Wnt signaling pathway in psoriasis. Conclusions ISG15 expression is increased in psoriatic cells and skin lesions of patients with psoriasis. In psoriasis, ISG15 promotes keratinocyte proliferation through the Wnt signaling pathway.
- psoriasis /
- keratinocyte proliferation /
- ISG15 /
- Wnt signaling pathway

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Figure 1. GO and KEGG enrichment analyses of differentially expressed genes derived fromGSE13355 and GSE30999 datasets

(A) Volcano map showing differentially expressed genes in GSE13355 and GSE30999 datasets. Orange color indicates high expression and blue low expression. (B) Venn diagram of the common differentially expressed genes (cross area) of GSE13355 and GSE30999 datasets. (C) Bubble map illustrating GO analysis (including biological process, cell component, and molecular function) and KEGG enrichment analysis of differentially expressed genes.

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Figure 2. Protein-protein interaction (PPI) networks of differentially expressed genes and real-time PCR quantification of expression levels of key genes

(A) PPI networks of differentially expressed genes. The size of the circle is proportional to the value. (B) Real time-PCR quantification of the expression levels of ISG15, LTF, and RSAD2 genes in psoriasis patients. ^**P < 0.01.

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Figure 3. Immunofluorescence staining of expression and distribution of ISG15 in psoriasis lesions

(A) Immunofluorescence (IF) was used to visualize the expression and distribution of ISG15 in psoriatic lesions. (B) Statistical data of immunofluorescent intensity. ^**P < 0.01.

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Figure 4. Silencing ISG15 in a psoriatic cell model and verifying its phenotypic proliferation efficiency

(A) Western blot images showing ISG15 protein expression after transfection with NC-siRNA or siISG15. (B) Statistical data of western blot bands. Note that silencing ISG15 reduced psoriatic cell proliferation. (C) Psoriatic cell proliferation rate following ISG15 knockdown determined by the MTT assays conducted on 1, 2, 3, 4 and 5 days after siISG15 transfection. ^**P < 0.01.

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Figure 5. Silencing ISG15 reduces the expression of Wnt signaling and regulates keratinocyte proliferation in psoriasis

(A) Western blot images depicting the changes of protein levels of β-catenin post-ISG15 silence. (B) Statistical data of Western blot bands for β-catenin. (C) Psoriatic cell proliferation rate with ISG15 knockdown was measured by MTT assay at 1, 2, 3, 4 and 5 days. (D) Western blot images showing the alterations of protein levels of β-catenin and Cyclin D1, a key regulator of cell cycle. (E-F) Statistics of Western blot bands. ^**P < 0.01.

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