Using bioinformatics approaches and experimental verification to identify HOXD8 as a key gene for keloid progression

Keloid is a cutaneous fibrotic disease characterized through excessive proliferation of fibrous tissue, which can induce scar swelling, pain and functional impairment. A full understanding of what causes keloid formation is still missing, and there aren't many effective treatment strategies to rely on. This study sought to explore the mechanisms of keloid, to offer novel targets for disease prevention and treatment. Gene Expression Omnibus (GEO) database was interrogated to select differentially expressed genes (DEGs) in GSE145725. PPI network analysis, gene enrichment analysis, and cell-specific and tissue-specific expression analyses of the hub gene were performed. To verify this hub gene, its expression of hub gene in normal skin tissue, pathological scar tissue and fibroblasts was analyzed by RT-qPCR and Western blot (WB). Small interfering RNA (siRNA) that specifically target hub gene were transfected into keloid fibroblasts. The expression levels of hub gene were detected by RT-qPCR and WB. CCK8, EDU, Transwell and Flow cytometry were used to evaluate cell proliferation, migration and apoptosis. Furthermore, changes in the Wnt/β-catenin signaling pathway were detected by WB. Bioinformatics analysis, revealed that Homeobox D8 (HOXD8) is a key gene in keloid. HOXD8 expression showed a significant increase of about 2-fold in keloid tissues. Silencing HOXD8 significantly reduced the proliferation and migration abilities of keloid fibroblasts, increased the apoptosis rate in association with the Wnt/β-catenin pathway. Our study demonstrated that HOXD8 may promote keloid formation via the Wnt/β-catenin signaling pathway in keloid fibroblasts. These findings improve our understanding of the molecular mechanisms underlying keloid pathogenesis and suggest HOXD8 as a potentially promising target for keloid intervention.