The effects of plasma from patients with active thyroid-associated orbitopathy on the survival and inflammation of melanoma-associated fibroblasts

• Published in: PeerJ (San Francisco, CA) CA), 2024-11, Vol.12, p.e18612, Article e18612
• Main Authors: Chen, Huifang, Chen, Shiyuan, Liu, Zhenfeng
• Format: Article
• Language: English
• Subjects: 1-Phosphatidylinositol 3-kinase; Adult; AKT protein; Antibodies; Cancer-Associated Fibroblasts - metabolism; Cancer-Associated Fibroblasts - pathology; Cell adhesion & migration; Cell Biology; Cell culture; Cell differentiation; Cell Movement; Cell Survival; Cholecystokinin; Coculture Techniques; Cytokines; Cytokines - blood; Cytokines - metabolism; Enzyme-linked immunosorbent assay; Ethylenediaminetetraacetic acid; Extracellular matrix; Extracellular signal-regulated kinase; Female; Fibroblasts; Fibroblasts - metabolism; Gene expression; Graves Ophthalmopathy - blood; Graves Ophthalmopathy - metabolism; Graves Ophthalmopathy - pathology; Humans; Immunofluorescence; Immunology; Inflammation; Inflammation - blood; Inflammation - metabolism; Invoices; Leukocyte migration; Macrophages; Male; Melanoma; Melanoma - blood; Melanoma - mortality; Melanoma - pathology; Melanoma-associated fibroblasts; Metastasis; Middle Aged; Oncology; Patient outcomes; Phosphatidylinositol 3-Kinases - metabolism; Phosphorylation; Plasma; Plasma - metabolism; Polarization; Reverse transcription; Signal Transduction; Skin cancer; Stat1 protein; Survival; Thyroid; Thyroid gland; Transcription activation; Tumor microenvironment; Tumorigenesis; Tumors; Vimentin
• ISSN: 2167-8359; 2167-8359
• DOI: 10.7717/peerj.18612

Plasma from patients with active thyroid-associated orbitopathy (TAO-A) could cause inflammation to fibroblasts, and such a mechanism was explored in the context of melanoma. Plasma samples collected from TAO-A patients and healthy control (HC) were primarily co-cultured with the melanoma-associated fibroblasts (MAFs) derived from melanoma patients. The survival and inflammation of the co-cultured MAFs were measured after confirming the levels of pro-inflammatory cytokines. Ki67 and Vimentin (VIM) markers were analyzed by immunofluorescence, and cell survival and migration were assessed using cell counting kit-8 (CCK-8) and Transwell. The THP-1 cells were induced to differentiate into macrophages, which were subsequently co-cultured to assess M1/M2 polarization status. Meanwhile, the levels of inflammatory factor were detected by enzyme-linked immunosorbent assay (ELISA). The gene expression was measured by reverse transcription quantitative PCR (RT-qPCR), and the activation of PI3K/AKT, STAT1, p65, and ERK signaling pathways was detected by Western Blot. Plasmas derived from TAO-A patients were characterized by elevated levels of pro-inflammatory cytokines, which enhanced the inflammation status and survival of MAFs, promoted the levels of PI3K and AKT, and downregulated expression of Bax. The co-culture of the plasma with MAFs evidently promoted M1 polarization and the phosphorylation of STAT1, P65 and ERK1/2. These findings proved the effects of the plasmas of TAO-A patients on the survival and inflammation of MAFs, providing evidence for future studies to delve into the relevant mechanisms.