Upregulation of Raf and Ralgds Gene Expression by Tgf-Β in Systemic Sclerosis Dermal Fibroblasts: A Controlled in Vitro Study Publisher Pubmed



Bakhshi F ; Mahalleh M ; Kavosi H ; Jafarisavari Z ; Ahmadzadeh N ; Enayati S ; Madreseh E ; Robatjazi B ; Mahmoudi M ; Farhadi E ; Vodjgani M
Source: Clinical Rheumatology Published:2025

Abstract

Introduction: Systemic sclerosis is an autoimmune rheumatic disorder characterized by uncontrolled fibroblast activation, skin thickening, and fibrosis. Activated Ras and ERK signaling significantly affect fibrosis and EMT upon TGF-β treatment. RalGDS may play an important role in inflammatory and oncogenic processes. This study investigates the expression of RAF and RalGDS genes in SSc patients compared to healthy individuals before and after treatment with TGF-β. Methods: This research included 20 patients with systemic sclerosis and 18 healthy controls matched for age and sex. Skin biopsies were collected, and fibroblasts were grown and expanded. The identity of fibroblasts was confirmed using immunofluorescence labeling. Fibroblasts were subjected to TGF-β1 treatment, and the expression levels of A-RAF, B-RAF, C-RAF, and RalGDS genes were assessed via real-time PCR. Results: Gene expression analysis revealed no significant baseline changes in RAF and RalGDS between groups; however, TGF-β therapy dramatically raised both genes in SSc cells relative to controls. A positive association was identified between the expression levels of the RAF and RalGDS genes. Conclusions: This study investigated A-RAF, B-RAF, C-RAF, and RalGDS gene expression in fibroblasts from SSc patients and healthy controls, focusing on their response to TGF-β treatment. While baseline expression levels were similar between groups, TGF-β significantly upregulated these genes in SSc fibroblasts more than in controls. This suggests that TGF-β signaling may contribute to the dysregulation of RAF and RalGDS pathways in SSc, highlighting their potential as biomarkers and therapeutic targets for the disease. (Table presented.) © 2025 Elsevier B.V., All rights reserved.