Unraveling the Dual Role of circ‐ CBLB and ETS ‐1 in Rheumatoid Arthritis: Biomarkers and Therapeutic Targets
Year: 2025
Type: article
Abstract: OBJECTIVE: To investigate the effects of circ-CBLB and ETS-1 on the proliferation, apoptosis, and inflammatory cytokine expression in the fibroblast-like synoviocytes (FLSs) from patients with rheumatoid arthritis (RA). METHODS: Peripheral blood was collected from 15 pairs of healthy controls (HCs) and patients with RA to isolate peripheral blood mononuclear cells (PBMCs). mRNA expression of circ-CBLB and ETS-1 was determined using qRT-PCR. Levels of the inflammatory markers (ESR, CRP, CCP, and RF) were determined, and the 28-joint Disease Activity Score (DAS28) was calculated. For in vitro experiments, human FLS and RA-FLS were cultured, and constructs (pcDNA3.1/siRNA-circ-CBLB, pcDNA3.1/siRNA-ETS-1) were transfected into RA-FLS. Cotransfection of pcDNA3.1-circ-CBLB and siRNA-ETS-1 was undertaken to explore their combined effects. Levels of the key inflammatory cytokines (interleukin [IL]-4, IL-23, IL-13, and tumor necrosis factor [TNF]-α) were evaluated using qRT-PCR and enzyme-linked immunosorbent assays. Functional assays (CCK-8) were used to assess cell viability, apoptosis (flow cytometry), and migration. Western blotting was used to determine protein expression. RESULTS: In vivo analysis showed significant downregulation of circ-CBLB and ETS-1 in PBMCs from patients with RA compared with the HCs, as confirmed using qRT-PCR. Correlation analysis indicated a positive association among circ-CBLB, ETS-1, and IL-4, while circ-CBLB and ETS-1 were negatively correlated with inflammatory markers (ESR, CRP, RF, CCP, DAS28, IL-23, and TNF-α). Receiver operating characteristic curve analysis suggested circ-CBLB and ETS-1 as potential biomarkers for high disease activity in RA. In vitro, circ-CBLB overexpression increased IL-4 levels while decreasing IL-23, IL-13, and TNF-α levels. Additionally, circ-CBLB inhibited the apoptosis of RA-FLS, prolonged the cell cycle, and reduced cell migration. ETS-1 negatively regulated circ-CBLB, indicating a feedback loop. CONCLUSION: circ-CBLB and ETS-1 are downregulated in RA and correlate with inflammation and disease activity. They regulate each other bidirectionally. circ-CBLB reduces RA-FLS viability, promotes apoptosis, and inhibits migration by modulating cytokines. ETS-1 has similar effects, and interfering with its expression reverses the impact of circ-CBLB.
Authors: Shu Li, HaoXiang Fang, Lei Wan, XiaoJun Zhang
Institution: Anhui University of Traditional Chinese Medicine
Language: English
Topic: Circular RNAs in diseases
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: bronze
Awards: G6705366635, G7922710459
OpenAlex ID: W4408510747