Abstract
Purpose:
We identified the mechanism and effects of chondrocyte-derived extracellular matrix (CDECM) on human pterygium epithelial cells (hPECs) migration.
Methods:
hPECs were isolated and cultured from specimens during surgical removal. The effects of CDECM (1mg/mL) on hPEC migration were evaluated according to a scratch wound closure assay, and trans-well invasion assay. In addition, to identified the ameliorative mechanism of CDECM on hPEC migration, we performed western blotting for expression of cytoskeletal proteins, MMPs, TIMPs and signaling molecules. The gleatinoytic activies of MMP-2 and MMP-9 were measured using gelatin zymography. The study was approved by the institutional review board (IRB) of Busan Paik Hospital, Inje University College of Medicine, Busan, Korea, approved the protocol (IRB No. 12128).
Results:
CDECM gradually inhibited cell migration and invasion. The migration of hPECs for 24 h was markedly increased. However, CDECM significantly inhibited to 0.34±0.08 fold in comparison with the control (p<0.05). CDECM also suppressed expression of cytoskeletal proteins including collagen Iα and fibronectin in hPECs. Furthermore, CDECM significantly inhibited MMP-2 and MMP-9 expression and activity, as well as increased TIMP-1 and TIMP-2 expression. In addition, CDECM markedly decreased expression of nucleus NF-κB p65 and Klf-4 through blocking the phosphorylation of p38MAPK, PKCα (Ser 657) and PKCθ (Thr538).
Conclusions:
Our findings suggest that the mechanism of suppression of the hPECs migration by CDECM resulted from the blocking the p38MAPK and PKC signaling pathways.
Keywords: 665 pterygium •
519 extracellular matrix