Abstract
Introduction: Stem cells fate and cellular functions are controlled by gene transcription, influenced by histone acetylation and deacetylation. Suberoylanilide hydroxamic acid (SAHA) is a clinically available histone deacetylase inhibitor. This study aim to understand SAHA’s role in stem cells regulation and its potential targets for tissue regeneration, including skin repair.Methods: Adipose-derived stem cells (ADSCs) treated with SAHA for 72 hours. Cell viability was evaluated using MTT assay. Western blotting was conducted to assess the expression of histone deacetylases (HDACs) and proliferating cell nuclear antigen (PCNA). Ki-67, a marker of cellular proliferation, was analyzed by immunofluorescence staining. Cell cycle progression was screened using propidium iodide staining. Differential gene expressions were identified through microarray analysis and validated using qPCR. Proteomic profiling was performed using mass spectrometry.
Results: Cell viability of ADSCs which treated with SAHA for 72 h, showed no change at the studied concentrations. However, the expression of histone deacetylase (HDACs) decreased at 1000 nM, while the cell proliferation marker Ki-67 increased after SAHA treatment. The expression of CCND1 gene expression was increased, and the proliferating cell nuclear antigen (PCNA) protein expression was decreased. Cell cycle analysis showed an increase in G2 phase. Microarray analysis revealed 74 upregulated and 40 downregulated differentially expressed genes, P53 targets, CDKN1A and MDM2 were among the upregulated genes. Proteomic analysis identified 631 upregulated and 823 downregulated proteins. Pathway enrichment analysis showed that cell cycle, ATP-dependent chromatin remodeling and DNA processes were the regulated pathways under the effect of SAHA.
Conclusion: This study suggests SAHA might alter ADSCs’ biological processes through coordinated manner which highlights its potential for skin regeneration.