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RHOA inhibits chondrogenic differentiation of mesenchymal stem cells in adolescent idiopathic scoliosis.

Authors :
Yang M
Chen K
Hou C
Yang Y
Zhai X
Chen K
Wei X
Bai Y
Li M
Source :
Connective tissue research [Connect Tissue Res] 2022 Sep; Vol. 63 (5), pp. 475-484. Date of Electronic Publication: 2022 Jan 12.
Publication Year :
2022

Abstract

Purpose: The etiology of adolescent idiopathic scoliosis (AIS) remains unclear. The chondrogenic differentiation of mesenchymal stem cells (MSCs) is important in AIS, and the Ras homolog gene family member A (RHOA) is associated with chondrogenesis. The purpose of this study was to explore the effect of RHOA on the chondrogenic differentiation of MSCs in AIS.<br />Methods: We isolated MSCs from patients with AIS (AIS MSCs) and individuals without AIS (control MSCs). The inhibitor Y27632 was used to inhibit the function of RHOA/ROCK signaling, and plasmid-based overexpression and siRNA-mediated knockdown were used to manipulate RHOA expression. CCK-8 was used to detect cell viability. The phosphorylation levels of LIMK1, MLC2 and cofilin were detected by Western blotting. The mRNA expression of aggrecan, SOX9, and COL2A1 were confirmed using RT-PCR. Immunofluorescence was used to analyze F-actin and collagen II. Alcian blue staining was performed to assess the secretion of glycosaminoglycans (GAGs).<br />Results: We found that RHOA was significantly upregulated in AIS MSCs, and the phosphorylation levels of LIMK1, MLC2, and cofilin were increased. The mRNA expressions of aggrecan, SOX9, and COL2A1 were notably reduced in AIS MSCs. However, these effects were abolished by Y27632 treatment and RHOA knockdown in AIS MSCs. In addition, RHOA knockdown in AIS MSCs increased the content of collagen II and GAGs. RHOA overexpression in the control MSCs markedly activated the RHOA/ROCK signaling and decreased the expression of aggrecan, SOX9, and COL2A1, F-actin, and GAGs.<br />Conclusion: RHOA regulates the chondrogenic differentiation ability of MSCs in AIS via the RHOA/ROCK signaling pathway and this regulation may involve SOX9.

Details

Language :
English
ISSN :
1607-8438
Volume :
63
Issue :
5
Database :
MEDLINE
Journal :
Connective tissue research
Publication Type :
Academic Journal
Accession number :
35019797
Full Text :
https://doi.org/10.1080/03008207.2021.2019247