Decorin knockdown affects the gene expression profile of adhesion, growth and extracellular matrix metabolism in C-28/I2 chondrocytes.
Journal
PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081
Informations de publication
Date de publication:
2020
2020
Historique:
received:
02
01
2020
accepted:
12
04
2020
entrez:
1
5
2020
pubmed:
1
5
2020
medline:
28
7
2020
Statut:
epublish
Résumé
Decorin is a member of small leucine-rich proteoglycan family, which is involved in multiple biological functions mainly as a structural and signaling molecule, and disturbances in its own metabolism plays a crucial role in the pathogenesis of osteoarthropathy. In this study, we aim to further explore the biological function of decorin and their role in human chondrocyte cell line, C28/I2. Lentivirus-mediated shRNA was applied to down-regulate decorin expression in C28/I2 chondrocytes. Effect of decorin knockdown on gene expression profiles was determined by RNA sequencing followed by bioinformatics analysis. MTT, adhesion assays and flow cytometry were used to investigate the effect of decorin knockdown on cell proliferation, adhesion, and apoptosis. sGAG content in the culture medium was determined by DMMB assay. Stably transfected C28/I2 cells were seeded onto the cancellous bone matrix gelatin (BMG) to construct tissue-engineered cartilage. The histological patterns were evaluated by H&E and Toluidine blue staining. In this study, 1780 differentially expressed genes (DEGs) including 864 up-regulated and 916 down-regulated genes were identified using RNA-Seq. The reliability of the gene expression was further verified by qRT-PCR. GO and KEGG pathway enrichment analysis revealed diverse cellular processes were affected by decorin silencing such as: cell adhesion, growth, and metabolism of extracellular matrix. In addition, we confirmed that down-regulation of decorin significantly suppressed cell proliferation and adhesion and induced apoptosis. The sGAG content in the media was significantly increased after decorin silencing. Engineered articular tissues in the decorin knockdown group exhibited cartilage destruction and proteoglycan loss as evidenced by H&E and Toluidine blue stains. Overall, this combined data helps to provide a comprehensive understanding of the roles of decorin following its knockdown in C28/I2 cells.
Identifiants
pubmed: 32353084
doi: 10.1371/journal.pone.0232321
pii: PONE-D-19-35519
pmc: PMC7192450
doi:
Substances chimiques
Decorin
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0232321Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
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