Effects of COL8A1 on the proliferation of muscle‐derived satellite cells

Collagen type VIII alpha 1 chain (COL8A1) is a component of the extracellular matrix. Our previous studies suggested that COL8A1 is associated with the proliferation of muscle‐derived satellite cells (MDSCs). Additionally, it has been demonstrated that COL8A1 promotes the proliferation of smooth mus...

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Bibliographic Details
Published in:Cell biology international 2018-09, Vol.42 (9), p.1132-1140
Main Authors: Li, Xiaofan, Wang, Zhao, Tong, Huili, Yan, Yunqin, Li, Shufeng
Format: Article
Language:English
Subjects:
AKT protein
Cell proliferation
COL8A1
Collagen
CRISPR
Cyclin B1
EGR-1 protein
EGR1
Enzyme inhibitors
Extracellular matrix
Hepatocytes
Kinases
Liver cancer
muscle‐derived satellite cell
PI3 K/AKT
proliferation
Protein kinase inhibitors
Satellite cells
Signal transduction
Smooth muscle
Western blotting
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Summary:Collagen type VIII alpha 1 chain (COL8A1) is a component of the extracellular matrix. Our previous studies suggested that COL8A1 is associated with the proliferation of muscle‐derived satellite cells (MDSCs). Additionally, it has been demonstrated that COL8A1 promotes the proliferation of smooth muscle cells and liver cancer cells. Therefore, we predicted that COL8A1 is associated with the proliferation of bovine MDSCs, which have potential applications in research. In this study, we constructed vectors to activate and repress COL8A1 in bovine MDSCs using the CRISPR/Cas9 technique and determined the effects of COL8A1 modulation by EdU labeling, Western blotting, and dual‐luciferase reporter assays. The results showed that activation of COL8A1 increased the number of EdU‐positive cells and expression of the proliferation markers cyclin B1 (CCNB1) and P‐AKT. The expression of P‐Akt was unchanged after addition of LY294002 (a protein kinase inhibitor capable of blocking the signal transduction pathway of the phosphoinositide 3‐kinase). In contrast, repression of COL8A1 reduced the number of EdU‐positive cells and expression of CCNB1 and P‐AKT. We also observed upregulation and downregulation of COL8A1 following the overexpression and repression of EGR1, respectively. The dual‐luciferase reporter assay revealed that EGR1 regulates the promoter activity of COL8A1. To our knowledge, this is the first study demonstrating that EGR1 positively regulates the expression of COL8A1, which in turn promotes the proliferation of bovine MDSCs via the PI3 K/AKT signaling pathway.
ISSN:1065-6995
1095-8355
DOI:10.1002/cbin.10979