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...

Full description

Saved in:
Bibliographic Details
Main Authors: Huifang Chen, Shiyuan Chen, Zhenfeng Liu
Format: Article
Language:English
Published: PeerJ Inc. 2024-11-01
Series:PeerJ
Subjects:
Online Access:https://peerj.com/articles/18612.pdf
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850221018437648384
author Huifang Chen
Shiyuan Chen
Zhenfeng Liu
author_facet Huifang Chen
Shiyuan Chen
Zhenfeng Liu
author_sort Huifang Chen
collection DOAJ
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
id doaj-art-6ed80cc44df049fc8b49f7c5a63c0271
institution OA Journals
issn 2167-8359
language English
publishDate 2024-11-01
publisher PeerJ Inc.
record_format Article
series PeerJ
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
work_keys_str_mv AT huifangchen theeffectsofplasmafrompatientswithactivethyroidassociatedorbitopathyonthesurvivalandinflammationofmelanomaassociatedfibroblasts
AT shiyuanchen theeffectsofplasmafrompatientswithactivethyroidassociatedorbitopathyonthesurvivalandinflammationofmelanomaassociatedfibroblasts
AT zhenfengliu theeffectsofplasmafrompatientswithactivethyroidassociatedorbitopathyonthesurvivalandinflammationofmelanomaassociatedfibroblasts
AT huifangchen effectsofplasmafrompatientswithactivethyroidassociatedorbitopathyonthesurvivalandinflammationofmelanomaassociatedfibroblasts
AT shiyuanchen effectsofplasmafrompatientswithactivethyroidassociatedorbitopathyonthesurvivalandinflammationofmelanomaassociatedfibroblasts
AT zhenfengliu effectsofplasmafrompatientswithactivethyroidassociatedorbitopathyonthesurvivalandinflammationofmelanomaassociatedfibroblasts