The effects of plasma from patients with active thyroid-associated orbitopathy on the survival and inflammation of melanoma-associated fibroblasts
Background 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. Methods Plasma samples collected from TAO-A patients and healthy control (HC) were primarily co-cultured with the...
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PeerJ Inc.
2024-11-01
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| author | Huifang Chen Shiyuan Chen Zhenfeng Liu |
| author_facet | Huifang Chen Shiyuan Chen Zhenfeng Liu |
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| description | Background 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. Methods 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. Results 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. Conclusion 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. |
| format | Article |
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| language | English |
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| spelling | doaj-art-6ed80cc44df049fc8b49f7c5a63c02712025-08-20T02:06:50ZengPeerJ Inc.PeerJ2167-83592024-11-0112e1861210.7717/peerj.18612The effects of plasma from patients with active thyroid-associated orbitopathy on the survival and inflammation of melanoma-associated fibroblastsHuifang Chen0Shiyuan Chen1Zhenfeng Liu2Department of Medicine, Guangxi University of Science and Technology, Liuzhou, ChinaDepartment of Traditional Chinese Medicine, The Second Affiliated Hospital of Guangxi University of Science and Technology, Liuzhou, ChinaDepartment of Traditional Chinese Medicine, The Second Affiliated Hospital of Guangxi University of Science and Technology, Liuzhou, ChinaBackground 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. Methods 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. Results 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. Conclusion 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.https://peerj.com/articles/18612.pdfMelanomaTumor microenvironmentMelanoma-associated fibroblastsInflammationSurvival |
| spellingShingle | Huifang Chen Shiyuan Chen Zhenfeng Liu The effects of plasma from patients with active thyroid-associated orbitopathy on the survival and inflammation of melanoma-associated fibroblasts PeerJ Melanoma Tumor microenvironment Melanoma-associated fibroblasts Inflammation Survival |
| title | The effects of plasma from patients with active thyroid-associated orbitopathy on the survival and inflammation of melanoma-associated fibroblasts |
| title_full | The effects of plasma from patients with active thyroid-associated orbitopathy on the survival and inflammation of melanoma-associated fibroblasts |
| title_fullStr | The effects of plasma from patients with active thyroid-associated orbitopathy on the survival and inflammation of melanoma-associated fibroblasts |
| title_full_unstemmed | The effects of plasma from patients with active thyroid-associated orbitopathy on the survival and inflammation of melanoma-associated fibroblasts |
| title_short | The effects of plasma from patients with active thyroid-associated orbitopathy on the survival and inflammation of melanoma-associated fibroblasts |
| title_sort | effects of plasma from patients with active thyroid associated orbitopathy on the survival and inflammation of melanoma associated fibroblasts |
| topic | Melanoma Tumor microenvironment Melanoma-associated fibroblasts Inflammation Survival |
| url | https://peerj.com/articles/18612.pdf |
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